Microcapsule composition for powdered probiotics and biocompounds based on double emulsions and method for producing same

Abstract

The present invention relates to a composition of microcapsules of powdered probiotics and biocompounds based on double emulsions, which includes a first emulsion of microorganisms dispersed in an oil phase with probiotic microorganisms or particles of same, wherein the oil phase contains biocompounds of plant or animal oil and a surfactant, and a second emulsion in aqueous phase with an oil phase corresponding to the first emulsion of microorganisms in oil phase, an aqueous phase with water and a biocompound ofa barrier material, wherein both phases are emulsified. Also described is a method for obtaining a composition of microcapsules of powdered probiotics and biocompounds based on double emulsions.

Description

[0001] Microcapsule composition for biocompounds and probiotic powders from double emulsions and method for preparing them

[0002] OBJECT OF THE INVENTION

[0003] The present invention may be included in the technical field of functional foods to supplement the diet. More specifically, the object of the present invention relates to microcapsule formulations for biocompounds and probiotic powders based on double emulsions that include ingredients that modulate the microbiota from said biocompounds.

[0004] BACKGROUND OF THE INVENTION

[0005] It has been found over the years that dysbiosis (imbalance of the intestinal microbiota) is related to different types of diseases in the body, from gastritis, inflammation of the intestines, colon cancer, to non-gastrointestinal diseases such as respiratory diseases, diabetes, obesity and cardiovascular diseases (Guerrant et al, 2008; Duvallet, et al., 2017; Turnbaugh et al. 2006). The study of this relationship between the alteration of the intestinal microbiota and different types of diseases has determined how certain bioactives or the absence of these can modify the intestinal microbiota, favoring the health of the organism that consumes these bioactives in the recommended quantities.An example of these bioactives are vitamins, which are micronutrients that promote the proper functioning of the body, keeping organs healthy, aiding energy collection, and the proper functioning of the immune system, among others (National Institute of Agriculture, 2019). Despite the beneficial effect of vitamins on the body, it is not capable of producing them on its own, so they must be acquired through the daily consumption of foods rich in these micronutrients (British Nutrition Foundation, 2019).

[0006] On the other hand, short-chain fatty acids are a product of intestinal microbiota fermentation. They are involved in regulating the immune system, maintaining the epithelial wall, and promoting a feeling of satiety after meals. Additionally, they are linked to the prevention of certain diseases, including diabetes, obesity, and colon cancer, among others (Baxter et al., 2019). To promote these fermentations, a diet rich in starch is necessary, the main substrate for the production of these short-chain fatty acids.

[0007] Over the years, it has been detailed that the occurrence of these fermentations and the production of these short-chain fatty acids are not only important for the function and prevention of diseases, but also because they intervene in the biosynthetic pathways of the psychological functions of the brain, including affective and cognitive processes (Dalile et al., 2019).

[0008] Additionally, probiotics are microorganisms that have a beneficial effect on health, and over the years, it has been shown that this beneficial effect arises from their modulatory effect on the gut microbiota. Their interaction with the gut microbiota leads to an increase in beneficial bacteria in the gut, as well as a reduction in the abundance of pathogenic species. This interaction could help restore the gut microbiota from dysbiosis, as discovered by Bo and colleagues (2020), where B. pseudolongum helped recover gut microbiota dysbiosis in obese mice, including microbiota diversity and the ratio of Firmicutes to Bacteroidetes. This treatment also increased the abundance of the bacterial genera Butyricimonas and Bifidobacterium. Singh et al., (2021 ) obtained similar results, in this study, old mice of general use were treated with Schizophyllum commune; this treatment significantly increased Bifidobacterium, Streptococcus and Lactobacillus, probiotic species of the intestinal microbiota.

[0009] Therefore, it is important to have an adequate intake of vitamins, healthy fatty acids and biotics (defined as probiotics, postbiotics, parabiotics or prebiotics), which can interact with the intestinal microbiota to prevent its imbalance and the malfunction of the system. One of the ways to protect these bioactives is by encapsulating them in such a way that the capsules protect them from the conditions of the gastrointestinal tract and are released in the intestine for interaction with the intestinal microbiota, thus being able to develop nutritional supplements that help satisfy the needs of the body and modulate the intestinal microbiota in favor of a balanced microbiota that achieves homeostasis. Thus, in the state of the art there are a plurality of disclosures related to microencapsulations of vegetable oils combined with probiotics, among these are:

[0010] Patent US11369644 teaches a consortium of bacteria of intestinal health-promoting species. This can be administered in a pharmaceutical composition in the form of drops, liquid, powder, or emulsion. The document does not refer to forms of protection or encapsulation of bacterial cells.

[0011] Similarly, patent application EP3609514 discloses a consortium of bacterial cells isolated from different sources. These cells are administered in a composition with liquid and powder pharmaceutical presentations. This consortium does not feature encapsulation for the cells.

[0012] For its part, patent document EP3930748 discloses a nutraceutical composition comprising prebiotics, probiotics and / or symbiotics, spirulina, cereals and micronutrients, which are microencapsulated. Specifically, spirulina is coated with GRAS lipids or beta-glucan and the bacterial probiotics are protected with whey proteins, natural polymer or alginate. This document does not assess the stability or release of these compounds in the food composition. Patent US9788563 discloses edible products, for example fruit juice-type beverages, containing encapsulated probiotic bacteria that are resistant to exposure to at least thermal and acidic conditions. The beverage products include at least one aqueous liquid and capsules comprising a gelled mixture of alginate and denatured protein, and probiotic bacteria trapped within the gelled mixture.The encapsulation process requires a mixture comprising sodium alginate, denatured protein, and active probiotic cells, and combining the mixture with a divalent cation to initiate cold gelation. It also requires a second mixture, which is filtered and encapsulated again.

[0013] Patent application WO2022160540 shows a method for preparing probiotic microcapsules by preparing a capsule core loaded with probiotics and preparing it in spherical particles using an extrusion-spheronization method; and coating these spheres by spray drying. This method, although it protects the probiotics, makes their preparation process more complex. The document by Eric-Parfait Kouamé, et.al (2023), which studies the encapsulation potential of probiotics (PRO) and linseed oil (FS) using different wall materials, . The addition of co-microcapsules also improved the amounts of polyunsaturated fatty acids (PUFA) released. However, it presents limited release of probiotics.

[0014] On the other hand, there is the document by Vimon, et.al. (2023) that reveals microcapsule networks based on polymeric matrices and charges under ionic crosslinking to encapsulate stable probiotics under adverse conditions. It also teaches the combination of agar (AG) and alginate (AL) through an o / w emulsion system for the incorporation of probiotics. The document mentions that by making a mixture of Lactobacillus plantarum MB001 (LPMB001) with AL and AG containing a few drops of soybean oil, a stable o / w microemulsion is obtained. However, the release of the components is irregular.

[0015] As can be seen, the state of the art requires improving encapsulation mechanisms to include components of diverse origin, both organic and non-organic, allowing microorganisms to remain stable and in homeostasis, and allowing their release to occur at the appropriate rate in the organism.

[0016] BRIEF DESCRIPTION OF THE INVENTION

[0017] This invention solves this problem by providing a microcapsule composition for biocompounds and probiotics. This composition is unique in that the compounds are presented in a powdered pharmaceutical form made from double emulsions that include ingredients that modulate the microbiota from the biocompounds.

[0018] The present invention relates to both a microcapsule composition for biocomposites and probiotics in powder form from double emulsions, and to the method for obtaining said composition.

[0019] In a first aspect, the invention relates to a microcapsule composition for biocomposites and probiotics in powder form from double emulsions that includes ingredients that modulate the microbiota, such as microorganisms of one or more genera, oil biocomposite and surfactants.

[0020] As mentioned above, said composition is composed of two emulsions, a first emulsion of particles dispersed in an oily phase that includes a biotic compound that can be a concentrated probiotic microorganism, probiotics, prebiotics, postbiotics or parabiotics and an oily phase that contains an oil, especially vegetable oil and a surfactant.

[0021] Said oily phase, initially obtained by emulsifying microorganisms dispersed in an oily phase containing a vegetable oil and a surfactant, is emulsified again in an aqueous phase containing water, a wall material, prebiotics and biocompounds.

[0022] Additionally, said composition in its oily phase includes lipophilic biocompounds solubilized in the oil, especially vegetable oil.

[0023] The invention also describes a method for obtaining said microcapsule composition for biocomposites and probiotics in powder form from double emulsions that includes the steps of solubilizing the microorganisms in the vegetable oil by means of simple mixing, mixing the surfactant and the vegetable oil with the solubilized microorganisms by means of simple mixing, and homogenizing.

[0024] Once the particles of a probiotic microorganism have been homogenized with the mixture of bio-compounds in the vegetable oil, they are homogenized using ultraturrax with the aqueous phase to obtain the formulation in liquid form.

[0025] The invention provides a double-emulsion microencapsulated composition containing probiotics and other biocompounds. This composition not only protects the probiotic content throughout the gastrointestinal tract but also reaches and interacts with the intestinal microbiota, positively modulating it. Furthermore, the presentation of the probiotic microorganisms in double-emulsion microcapsules allows them to remain stable, such that their viability is greater than in simple encapsulations or other types of pharmaceutical presentations. BRIEF DESCRIPTION OF THE FIGURES.

[0026] To complement the description being made and in order to help better understand the characteristics of the invention, in accordance with a preferred example of practical implementation thereof, a set of drawings is attached as an integral part of said description, in which the following has been represented for illustrative and non-limiting purposes:

[0027] Figure 1.- Digestion of 3 powdered microencapsulates. MC1: Microencapsulated probiotic and lipophilic biocompounds in high oleic palm oil; MC2: Microencapsulated probiotic and lipophilic biocompounds in high oleic palm oil, with hydrophilic postbiotics produced by probiotic fermentation; MC3: Microencapsulated probiotic and lipophilic biocompounds in high oleic palm oil, with hydrophilic prebiotics.

[0028] Figure 2. Relative abundance of the top 10 most abundant species. G5 and G6 before treatment with MC1. G3 and G4 after treatment with MC1.

[0029] Figure 3. Relative abundance of the top 10 most abundant genera. G5 and G6 before treatment with MC1. G3 and G4 after treatment with MC1.

[0030] DETAILED DESCRIPTION OF THE INVENTION

[0031] A preferred embodiment of the object of the invention is described below with the help of figures.

[0032] In a first aspect of the invention, a microcapsule composition for biocomposites and probiotics in powder form from double emulsions is described, which includes ingredients that modulate the microbiota from bio-composites.

[0033] Said composition includes a first emulsion of particles dispersed in an oily phase containing a probiotic microorganism, preferably concentrated, which is between 1 and 50%, preferably between 20 and 40% and more preferably between 36.3% and 38.8% w / w of the first emulsion. This microorganism can be selected from Lactobacillus, Bacillus, Limosilactobacillus, Bifidobacterium, Streptococcus, Enterococcus and other genera of microorganisms known for their probiotic function.

[0034] Among these microorganisms, Lactobacillus are preferably selected from: Lactobacillus delbrukii, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus rhamnosus, and mixtures thereof. Likewise, among Enterococcus, Enterococcus faecium or another microorganism recognized as a probiotic is preferable. However, the composition is not limited to these species.

[0035] It should be understood that, in accordance with the nature of the invention, restricting it to a single probiotic microorganism is not appropriate since the central objective of this is to obtain powdered microcapsules from double emulsions that include ingredients that modulate the microbiota and therefore the invention is applicable to any type of probiotic microorganism, as well as to any biocompound.

[0036] Said composition further comprises an oily phase containing an oil, which can be selected from vegetable oil or oil of animal origin. The vegetable oil is selected from: coconut, corn, chia, canola, fish, argan, sesame, hazelnut, sunflower, olive, palm, linseed and high oleic oils, rich in individual omega 3 and 6, or a mixture thereof. This oil(s) constitute(s) between 30 and 70%, preferably between 50 and 70% and more preferably between 58.2% and 60.6% w / w of the oily phase. In addition, there is a surfactant corresponding to soy lecithin or polyoxyethylene (20) sorbitan monooleate (Tween 80) and which constitutes between 1 and 10%, preferably between 1 and 4% and more preferably between 2.9% and 3.03% w / w of the oil phase.

[0037] The composition further includes a second emulsion containing the oily phase obtained previously between 1 and 10% w / w, preferably between 5.15 and 8.25% w / w of the final emulsion (the oily phase containing probiotic microorganisms, the oil and the surfactant) and an aqueous phase containing water which is between 40 and 70% w / w, preferably between 49.6% and 64.4% w / w of the aqueous phase, a wall material selected from soy protein, ovalbumin, alpha-lactoglobulin, albumin immunoglobulin, zein; and starches such as potato starch, rice, maltodextrin, cassava which is between 20% and 40%, preferably between 24.1% and 35.5% w / w of the aqueous phase. In one embodiment of the invention the aqueous phase includes a prebiotic that is selected from oligosaccharides, xylooligosaccharides, fructooligosaccharides, which are found between 1% to 2% w / w of the aqueous phase;

[0038] In another embodiment of the invention, the aqueous phase includes biocompounds selected from vitamins, such as vitamin C and B complex, and minerals, such as iron, calcium, zinc, among others of a similar nature, and which are found between 1% and 2% w / w of the aqueous phase; where the oily phase containing particles of a concentrated probiotic microorganism, the vegetable oil and the surfactant, and an aqueous phase are emulsified.

[0039] In another embodiment of the invention, the microorganism that is in particle form is either alive or dead.

[0040] In another embodiment of the invention, the aqueous phase of the composition includes a supernatant where the microorganism was cultured and which is between 30% and 35% of the final emulsion.

[0041] In a further embodiment of the invention, the aqueous phase includes lipophilic bio-compounds solubilized in the vegetable oil and which constitutes between 36.3% and 38.8% of the oily phase, where the lipophilic bio-compounds are selected from lipophilic vitamins such as B, E, A, D, medium and short chain fatty acids and are found in 1.82% of the final emulsion.

[0042] In another aspect of the invention, a method is described for obtaining a microcapsule composition for biocomposites and probiotics in powder form from double emulsions that includes ingredients that modulate microbiota from bio-composites, comprising the steps of:

[0043] • solubilize lipophilic bio-compounds in the oil, preferably vegetable oil, by means of simple mixing;

[0044] • After solubilization, the surfactant and vegetable oil should be mixed with the solubilized lipophilic biocompounds using a simple mixture. • After these initial mixtures are solubilized, the probiotic microorganisms are added and homogenized with Ultraturrax at a speed between 5000 and 8000 rpm, for a time between 2 and 8 minutes,

[0045] • After having this mixture homogenized, the previously defined aqueous phase is added and homogenized with Ultraturrax between 8000 and 15000 rpm, for a time between 2 and 8 minutes.

[0046] In another optional embodiment of the invention, the described method also includes the optional step of drying the emulsion obtained in the last described homogenization by means of spray drying with an inlet temperature between 150 and 210 °C, an outlet temperature between 80 and 90 °C and an atomization pressure between 1.48 and 2.467 atm.

[0047] This process allows the probiotic microorganisms to be encapsulated initially by the oil that makes up the oil phase and additionally by the wall material of the aqueous phase in a double emulsion. When this double emulsion is spray-dried, most of the water in the emulsion is removed, leaving the probiotic protected by the oil and the powdered wall material.

[0048] In this way, the probiotic is given such protection that the thermal processes to which it may be subjected when incorporated into solid food matrices do not diminish its viability, and likewise, when it passes through the gastrointestinal tract, whose varying pH and salt conditions and its interaction with its own enzymes, do not affect its viability and structure, thus arriving in a concentration adequate for interaction with the intestinal microbiota and exerting its modulating effect on it.

[0049] In a further embodiment, the invention describes a method comprising the optional step of vacuum packing the microcapsule powder obtained after spray drying.

[0050] In order to show the advantages and technical characteristics of the invention, a non-limiting example of the method described in the present invention, in vitro digestion tests of the composition described in the present invention and in vitro fermentation tests of the composition with intestinal microbiota are shown below. Examples

[0051] Example 1. Preparation of the double emulsion P / O / W (Particle in oil in water)

[0052] High oleic palm oil in the proportions indicated in the experimental design (Table 1) is mixed with soy lecithin as a surfactant in a proportion of 3% with respect to the amount of oil used. The concentrated biotic (Probiotic L fermentum) is slowly added to the oil and homogenized at 8000 rpm in an Ultraturrax N25 (IKA, US) for 2 min. Separately, 11% whey as a protein emulsifier is added to the water in the proportions indicated by the design and mixed at 11000 rpm for 8 min. Subsequently, the oily phase composed of the oil and the biotic is slowly added to the aqueous phase at 11000 rpm for 5 min, obtaining the double emulsion P / O / W. This emulsion is subjected to spray drying at an inlet temperature of 200 °C and an outlet temperature of 90 °C. The atomization pressure varies according to the experimental design presented in Table 1.

[0053] After obtaining the powdered microencapsulates, 1 g of these were reconstituted at a 10% ratio in 0.1% peptone water and seeded at different dilutions on MRS agar to observe viability after drying. Additionally, 1 g of the obtained powders were placed in contact with 9 mL of 0.3% porcine bile for 2 h to observe the viability of the probiotic microencapsulate after contact with bile. The results are presented in Table 1.

[0054] Table 1. Design of experiments to obtain a microencapsulated probiotic and biocomposites.

[0055] Probiotic Run Oil Serum Pressure Difference in Difference in Percentage of (bar) cycles log cycles log CFU / g moisture (%H)

[0056] (% w / w) (% w / w) (% w / w) UFC / g

[0057] 1 7.67 12.00 20.33 1 .50 1 .7 2.5 1 .20

[0058] 2 8.00 12.00 20.00 2.00 2.4 1 .8 3.85

[0059] 3 2,60 4,07 33,33 2,00 1 ,6 1 ,5 2,44

[0060] 4 7,67 12,00 20,33 1 ,50 1 ,8 2,8 1 ,44

[0061] 5 5,85 9,15 20,00 2,00 0,9 1 ,2 2,84

[0062] 6 2,67 4,00 28,33 1 ,50 1 ,6 1 ,7 2,11

[0063] 7 2,67 4,00 28,33 2,00 1 ,2 0,9 1 ,89

[0064] 8 3,75 5,71 26,99 1 ,63 0,3 3,3 2,46

[0065] 9 2,60 4,07 33,33 2,00 0,6 3,1 2,50

[0066] 10 6,25 9,71 22,99 1 ,63 1 ,0 2,1 2,41

[0067] 11 2,67 4,00 28,33 1 ,50 0,8 2,0 2,21

[0068] 12 4,82 7,42 25,66 1 ,75 1 ,5 1 ,8 2,69

[0069] 13 8,00 12,00 20,00 2,00 1 ,8 1 ,6 1 ,44

[0070] 14 5,85 9,15 20,00 2,00 0,5 3,6 4,13

[0071] 15 3,71 5,74 29,49 1 ,88 1 ,5 2,6 2,61

[0072] An analysis of variance was performed to observe how the variation of the 4 chosen factors influenced the percentage of powder moisture and the viability of the probiotic after contact with bile. A 2IF model was fitted to the behavior of these two response variables with R2 above 0.98 in both cases and a significance of p < 0.05. For the response variable of the difference in logarithmic cycles in probiotic viability after contact with bile, it was observed that individual factors and the interaction between the proportion of probiotic and oil with air affect the difference in logarithmic cycles. These same factors are also those that affect the percentage of moisture, allowing us to infer that the percentage of moisture and the viability of the probiotic are related. The equations for the prediction of each of these variables are presented below, based on the factors and the interaction between them:

[0073] Eq. 1.

[0074] Difference in cycles after bile = -7.05-175.52*A+147.61 *B- 11 .22*C+6.44*D-13.92*AB- 31 *AC+243.36*AD+14.82*BC-226.68*BD

[0075] Eq. 2.

[0076] % H = 5.01 + 38.74 *A-31.18*B+3.97*C-1.49*D+3.22*AB+15.07*AC-68.35*AD- 10.23*BC+64.25*BD

[0077] The response variables in this experimental design were optimized using response surface optimization, where the objective was to minimize both variables. The analysis between the theoretical optimum and the experimental optimum yielded a difference of no more than 5%.

[0078] Additionally, an alternative formulation with another oil described below was tested, in this case the olive oil was mixed with Polyoxyethylene (20) sorbitan monooleate (Tween80) (at 5% of the oil quantity) and with the probiotic L. plantarum in the formulation proportions of the optimum found previously. This mixture was homogenized at 8000 rpm in an Ultraturrax N25 (IKA, US) for 2 min. Separately, rice starch and guar gum as emulsifier are added to the water in the proportions indicated by optimum found with the previous design and mixed at 11000 rpm for 8 min. Subsequently the oily phase composed of the oil and the probiotic is slowly added to the aqueous phase at 11000 rpm for 5 min, obtaining the double emulsion P / O / W.This emulsion is subjected to spray drying at an inlet temperature of 180°C and an outlet temperature of 80°C, where the probiotic microorganism survived at 80%, similar behavior to that obtained with whey as the wall material and high oleic palm oil as the oil phase oil.

[0079] Example 2. In vitro digestion of microencapsulated biotics and powdered biocomposites.

[0080] The biotic and biocomposite microcapsules were subjected to in vitro digestion following the INFOGEST 1.0 protocol proposed by Minekus et al. (2014). In this case, the viability of the probiotic present in each of the microencapsulates was analyzed after each of the digestion phases (oral, gastric, and intestinal). The results are presented in Figure 1.

[0081] These results show how the microcapsules effectively protect the probiotic, allowing it to remain in high concentrations after the end of the intestinal phase. Under normal conditions, unencapsulated probiotics do not survive their passage through the gastrointestinal tract, since the conditions of digestion cause the microorganism to lose viability, reaching the intestine in minimal concentrations. Thus, the fact that it retains its viability throughout its passage through the gastrointestinal tract makes the protection provided by encapsulation effective.

[0082] Example 3. In vitro fermentation of microencapsulated biotics and biocomposites with intestinal microbiota of children with and without autism spectrum disorder (ASD).

[0083] After in vitro digestion, the MC1 microcapsule was subjected to in vitro fermentation with intestinal microbiota from a child with ASD and a child without ASD to observe changes in the colon's microbial structure. The same microbiota samples were used as controls for comparison, but without treatment with the microcapsule. In both cases, an enrichment of probiotic species was observed (Figure 2) compared to the controls.

[0084] It is possible to observe how different groups of beneficial species are enriched after treatment with MC1, such as Lactobacillus fermentum, Bifidobacterium longum, and Bifidobacterium adolescentis, in both the ASD microbiota samples and the microbiota of children without ASD. Additionally, genera such as Bifidobacterium and Limosilactobacillus increase after treatment in both types of samples, while genera such as Coprococcus and Enterococcus increase in samples from children without ASD, as shown in Figure 3.

[0085] The aforementioned species and genera that were enriched in the contact of the microbiota with the MC1 microcapsules are important for the correct development of intestinal homeostasis. Genera such as Bifidobacterium and Limosillactobacillus have probiotic strains such as Bifidobacterium lognum and Lactobacillus fermentum which help reduce the expression of proinflammatory cytokines, help balance intestinal immunity and repair and reinforce the intestinal mucosal barrier (Ehrlich et al., 2020), which in children with ASD has been shown to be decreased. On the other hand, the Coprococcus genus is associated with increased intestinal function, having a beneficial antagonistic role against pathogens, increasing the production of antimicrobial compounds by the intestinal microbiota that help against pathogens in the host (Yang et al., 2023).Additionally, the decrease in this Copprococcus genus has been found in patients suffering from depression (Suseelan et al., 2023), with its enrichment and balance being a good result in either of the two populations studied.

[0086] The previous examples show how the microcapsules obtained through the process and formulation proposed in the present invention protect the biotics and biocompounds from the conditions of the gastrointestinal tract and allow them to reach the intestinal microbiota in concentrations suitable for interaction, modulating its microbial structure by enriching genera and species that are beneficial for homeostasis and its balance, thus fulfilling its purpose.

[0087] Bibliographic references

[0088] Baxter, N. T., Schmidt, A. W., Venkataraman, A., Kim, K. S., Waldron, C., & Schmidt, T. M. (2019). Dynamics of human gut microbiota and short-chain fatty acids in response to dietary interventions with three fermentable fibers. MBio, 10(1 ).

[0089] Bo, T. bei, Wen, J., Zhao, Y. chun, Tian, S. jie, Zhang, X. ying , & Wang, D. hua. (2020). Bifidobacterium pseudoIongum reduces triglycerides by modulating gut microbiota in mice fed high-fat food. Journal of Steroid Biochemistry and Molecular Biology, 198(November 2019), 105602. https: / / doi.Org / 10.1016 / j.jsbmb.2020.105602

[0090] Dalile, B., Van Oudenhove, L., Vervliet, B., & Verbeke, K. (2019). The role of short-chain fatty acids in microbiota-gut-brain communication. In Nature Reviews Gastroenterology and Hepatology (Vol. 16, Issue 8, pp. 461 —478). Nature Publishing Group. https: / / doi.Org / 10.1038 / S41575-019-0157-3

[0091] Duvallet, C., Gibbons, S. M., Gurry, T., Irizarry, R. A., & Alm, E. J. (2017). Meta-analysis of gut microbiome studies identifies disease-specific and shared responses. Nature Communications, 8(1 ), 1-10. https: / / doi.org / 10.1038 / s41467-017-01973-8 Ehrlich AM, Pacheco AR, Henrick BM, Taft D, Xu G, Huda MN, Mishchuk D, Goodson ML, Slupsky C, Barile D, Lebrilla CB, Stephensen CB, Mills DA, Raybould HE. lndole-3- lactic acid associated with Bifidobacterium-dominated microbiota significantly decreases inflammation in intestinal epithelial cells. BMC Microbiol. 2020 Nov 23;20(1 ):357. doi: 10.1 186 / S12866-020-02023-y.

[0092] Guerrant, R. L., Oriá, R. B., Moore, S. R., Oriá, M. O. B., & Lima, A. A. A. M. (2008). Malnutrition as an enteric infectious disease with long-term effects on child development. In Nutrition Reviews (Vol. 66, Issue 9, pp. 487-505). NIH Public Access. https: / / doi.Org / 10.1111 / i.1753-4887.2008.00082.x

[0093] Kouadio Jean Eric-Parfait Kouamé, Awa Fanny Massounga Bora, Yibo Liu, Xiaoxue Yu, Yue Sun, Muhammad Hussain, Masumuzzaman Md, Ibourahema Coulibaly, Xiaodong Li, Lu Liu. Development and characterization of omega-3-rich flaxseed oil microcapsules and evaluation of its stability and release behavior in probiotic millet yogurt. Powder Technology, Volume 427, 2023,118739.

[0094] Minekus, M., Alminger, M., Alvito, P., Ballance, S., Bohn, T., Bourlieu, C., Carriere, F., Boutrou, R., Corredig, M., Dupont, D., Dufour, C., Egger, L., Golding, M., Karakaya, S., Kirkhus, B., Le Feunteun, S., Lesmes, U., Macierzanka, A., Mackie, A., ... Brodkorb, A. (2014). A standardised static in vitro digestion method suitable for food - an international consensus. Food Funct., 5(6), 11 13-1124. https: / / doi.org / 10.1039 / C3F060702J.

[0095] Ruipeng Yang, Shuhua Shan, Jiangying Shi, Hanging Li, Ning An, Songtao Li, Kaili Cui, Huiqin Guo, and Zhuoyu Li. Journal of Agricultural and Food Chemistry 2023 71 (7), 3273-3284. DOI: 10.1021 / acs.jafc.2c0669.

[0096] Sasi Vimon, Thanit Kertsomboon, Suwabun Chirachanchai, Kris Angkanaporn, Chackrit Nuengjamnong. Matrices-charges of agar-alginate crosslinked microcapsules via o / w microemulsion: A non-spore forming probiotic bacteria encapsulation system for extensive viability. Carbohydrate Polymers, Volume 321 , 2023,121302.

[0097] Singh, V., Muthuramalingam, K., Kim, Y. M., Park, S., Kim, S. H., Lee, J., Hyun, C., Unno, T., & Cho, M. (2021 ). Synbiotic supplementation with prebiotic Schizophyllum commune derived |3-(1 ,3 / 1 ,6)-glucan and probiotic concoction benefits gut microbiota and its associated metabolic activities. Applied Biological Chemistry, 64(1 ). https: / / doi.Org / 10.1 186 / s13765-020-00572-4

[0098] Suseelan, S., & Pinna, G. (2023). Heterogeneity in major depressive disorder: The need for biomarker-based personalized treatments. Advances in Clinical Chemistry, 112, 1 - 67. https: / / doi.org / 10.1016 / BS.ACC.2022.09.001 Turnbaugh, P. J., Ley, R. E., Mahowald, M. A., Magrini, V., Mardis, E. R., & Gordon, J. I. (2006). An obesity-associated gut microbiome with increased capacity for energy harvest. Nature, 444(7122), 1027-1031. https: / / doi.org / 10.1038 / nature05414.

Claims

CLAIMS 1. Composition of microcapsules of biocomposites and probiotics in powder form from double emulsions comprising: A. a first emulsion of microorganisms dispersed in an oily phase with: i. probiotic microorganisms or particles thereof between 20% and 50% w / w of the first emulsion, where the microorganisms are selected from: Lactobacillus, Bacillus, Limosilactobacillus, Bifidobacterium, Streptococcus, Enterococcus or mixtures thereof; where the oily phase contains: i. a biocompound of vegetable or animal oil constituting between 50% and 70% w / w of the oil phase; and iii. a surfactant selected from soy lecithin, polyoxyethylene(20)sorbitan monooleate, and constituting between 1% and 4% w / w of the oil phase; B. a second emulsion in aqueous phase with: i. an oil phase corresponding to the first emulsion of microorganisms in oil phase, according to stage A; between 2% and 10% i. an aqueous phase with: • water between 40 to 70% w / w of the aqueous phase; • A biocomposite of a wall material comprising 20 to 40% w / w of the aqueous phase, selected from: soy protein, ovalbumin, alpha-lactoglobulin, albumin immunoglobulin, zein; and starches such as potato starch, rice starch, maltodextrin, cassava starch, and mixtures thereof; where both phases are emulsified.

2. The composition according to claim 1 wherein the vegetable oil is selected from: coconut, corn, chia, canola, argan, sesame, skin oil Hazelnut, sunflower, palm, linseed and high oleic oils, rich in individual omega 3 and 6 or a mixture of these.

3. The composition according to claim 1 characterized in that the microorganism is alive or dead.

4. The composition according to claim 1 wherein the probiotic Lactobacillus microorganisms are selected from: Lactobacillus delbrukii, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus rhamnosus or mixtures thereof.

5. The composition according to claim 1 to 2, characterized in that the aqueous phase also includes a prebiotic that is selected from oligosaccharides, xylooligosaccharides, fructooligosaccharides and that is found between 1% and 2% of the aqueous phase.

6. The composition according to claim 1 to 3, characterized in that the aqueous phase also includes a biocompound that is selected from vitamins and minerals and that is found between 1% and 2% of the aqueous phase.

7. The composition according to claim 1 characterized in that the aqueous phase includes a supernatant where the microorganism was cultured and which is between 30% and 35% of the final emulsion.

8. The composition according to claim 1, characterized in that the aqueous phase includes lipophilic biocompounds solubilized in the vegetable oil and which constitutes between 36.3% and 38.8% of the oily phase, where the lipophilic biocompounds are selected from lipophilic vitamins such as B, E, A, D, medium and short chain fatty acids and are found in 1.82% of the final emulsion.

9. A method for obtaining a composition of microcapsules of biocomposites and powdered probiotics from double emulsions comprising: a. solubilizing biocompound in the oil by means of simple mixing; b. mix by a simple mixture of the surfactant and the oil with the biocompounds solubilized in step a); c. add the probiotic microorganism and homogenize with the mixture obtained in step b); d. mix the aqueous phase and the homogenized mixture in step c) and homogenize.

10. A method according to claim 9 comprising drying the emulsion in step d) by spray drying with an inlet temperature between 150 and 210 °C, an outlet temperature between 80 and 90 °C and an atomization pressure between 1.5 and 2.5 bar. 1 1. A method according to claim 10 comprising vacuum packing the microcapsule powder obtained after spray drying.

12. A method according to claim 9, characterized in that the homogenization of step c) is carried out with ultraturrax at a speed between 5000 and 8000 rpm, for a time between 2 to 8 minutes.

13. A method according to claim 9, characterized in that the homogenization of step d) is carried out with Ultraturrax between 8000 and 15000 rpm, for a time between 2 and 8 minutes.

Images