WO2019144979A1

WO2019144979A1 - Method for granulating and coating probiotics and granulated core obtained using same

Info

Publication number: WO2019144979A1
Application number: PCT/CO2018/000004
Authority: WO
Inventors: Sandra Janneth SANTOS ROCHA
Original assignee: Vallecilla B Y Vallecilla M Y Cia Sca Carval De Colombia
Priority date: 2018-01-25
Filing date: 2018-01-25
Publication date: 2019-08-01

Abstract

The invention relates to a method for coating and encapsulating probiotics, which comprises two steps: one of granulating and another of subsequently coating in a fluid bed dryer. The method allows longer and stable storage of that type of product without the need for refrigeration. The granulated core thus obtained ensures probiotic survival rates of more than 70% for up to 12 months, at an ambient temperature of up to 30 °C and without the need for refrigeration, and is suitable for inclusion in various food and pharmaceutical matrices. This technology combines two types of aspersion: a top spray for the granulation process, which is aimed at generating a granule with a suitable and functional size, and a subsequent tangential spray to optimise the process of coating the granule, ensuring the coverage thereof and the protection of the probiotics.

Description

PROCEDURE FOR GRANULATION AND COATING OF PROBIOTICS AND GRANULATED CORE OBTAINED THROUGH THE SAME

OBJECT OF THE INVENTION

The present invention belongs to the field of pharmaceutical chemistry and biotechnology. It reveals a method of coating and encapsulation for probiotics, which allows extended and stable storage of this kind of products without refrigeration.

BACKGROUND

Modern lifestyle habits and the use of uncontrolled antibiotics, has caused the balance of the gastrointestinal tract to be affected and increases the risk of developing diseases of all kinds, from gastrointestinal, to those related to the immune system.

Probiotics represent a prevention and treatment strategy for all these disorders. The clinical use of probiotics is gaining increasing attention worldwide, both in medical practice and in personal care in general (Masi, 2013), attending different gastrointestinal, urogenital, respiratory, oral and immunological conditions (Hathout et al , 2011).

According to the World Health Organization (FAO / WHO, 2001), probiotics are living microorganisms that, when ingested, promote host health. These microorganisms are considered normal flora of the gastrointestinal tract of animals and humans. Probiotics represent a potential in terms of health and nutritional benefits, since they control pathogenic microorganisms, providing protection to the host, whether human or animal (Hathout et al, 2011).

In humans, there is a growing trend towards a healthy and balanced diet, with natural ingredients that maintain consumer health. This is how probiotics are gaining more and more market in the field of dairy and nutritional supplements, without the medicine sector; where their virtues are proven (Market and Market, 2010). On an industrial level, these microorganisms are produced by fermentation technologies, which consists in the propagation of microbial cells in bioreactors, in order to obtain enough biomass and then be concentrated through separation and drying processes (Lacroix, 2007).

Being microorganisms living beings very sensitive to adverse oxygen conditions, high temperatures, humidity and pH, the processes of obtaining and formulating these probiotics are critical, since their performance will depend on the level of the gastrointestinal tract where they act . For this reason, some strategies have been developed at the level of fermentation process (Lacroix, 2007) or already in the drying process, using microencapsulation and coating technologies (Del Piano et al, 2011).

Microencapsulation is the process by which small particles or drops are surrounded by a coating agent producing microcapsules. There are different microencapsulation processes and different purposes. Depending on the application, the coating agents can vary from polysaccharides to lipids. The former have been used in aqueous solution applications such as yogurt and some beverages, while lipids have been applied to matrices such as creamy and related cheeses.

The technologies most commonly used in probiotic microencapsulation processes include extrusion, emulsion and drying.

The extrusion technology aims to develop beads or gel capsules that are made from hydrocolloids. Powdered probiotics are mixed with the hydrocolloid; then, said mixture is passed through an extruder generating micro drops that fall into a gelling solution under permanent agitation. The size of the microgranules will depend on the orifice of the extruder and the distance between it and the gelling solution. In the end you get a pearl with solid porous center. (Gbassi et al, 2012).

For its part, the emulsion process consists in the dispersion of the probiotic-hydrocolloid mixture in a vegetable oil. The resulting water-in-oil emulsion is homogenized by permanent agitation. This is the critical stage that generates the size and shape of the drops formed that are then collected by sedimentation. In the end, microcapsules with liquid center are obtained. (Gbassi et al, 2012).

As for drying technologies, lyophilisate, spray drying and bed drying are found. Lyophilization is a process in which water is removed from a frozen solution by sublimation under pressure. This generates dry products of good quality, without loss of sensory or nutritional characteristics. These products can be easily rehydrated for later use. This technology is very expensive due to the high energy expenditure and the duration of the process compared to other methods (Barbosa J, 2015)

For its part, spray drying allows the transformation of a solution into a dry powder in one step. Said solution is fed to a chamber where there is hot dry air and evaporates the water contained in the solution, generating a powder of good characteristics. This technology is cheaper than lyophilization and faster. (Barbosa J, 2015)

Drying of the fluid bed consists of liquid spraying in a bed of fluidized particles. The sprayed liquid is absorbed by an inert support or binding agent creating a core that is coated with layers. This occurs while hot air is injected that fluidizes the material and evaporates the liquids. The particles can grow by agglomeration and this is known as granulation. These processes are performed in batches. The resulting product can have different sizes depending on the desired one, since the variables can be modified to achieve it.

In extrusion and emulsification technologies, probiotic bacteria are enclosed in a gelatinous matrix of natural biological materials such as alginates, carrageenans or gums. The main difficulty is that due to its jelly-like consistency they can be stabilized in liquids only. These processes are not easily industrial ladders (Semyonov et al, 2012).

As for drying technologies, lyophilization and spray drying generate microcapsules; while fluid bed drying generates a microcapsule with a core and a true cover.

Freeze drying produces microcapsules of probiotics with good viability characteristics; However, it is expensive and its industrial escalation is difficult. Spray drying is widely used, but the temperature characteristics that are handled and the osmotic pressure exerted is so strong that cell viability is affected.

In the fluid bed drying process, the operating conditions further favor the viability and stability of the probiotic due to the temperatures it handles, not exceeding 40 ° C, it occurs in shorter times and is economically more viable. Similarly, sizes adjusted to the needs of subsequent application are obtained (Semyonov et al, 2012). In the end, what is sought with the different methods of encapsulation of probiotics is that these, once introduced to the application matrix, remain viable, retain their probiotic properties and maintain their stability over time, remaining in the final product in the desired quantities, until the end of the useful life of the finished product.

Similarly, some microencapsulation methods seek gastrointestinal resistance, which implies that the finished product requires refrigerated storage, a condition that makes storage, transport and final distribution of products more difficult.

All the methods explained partially fulfill these conditions and even allow the matrices encapsulated with probiotics to maintain their stability for a limited number of hours or days. However, none of these methods, even in combination with others, allows the transported probiotic to be kept within the final product in the desired quantities and during its useful life, which can usually be one to two years.

On the other hand, numerous studies and patents related to different probiotic microencapsulation techniques have been reported:

First, the patent document W02009 / 070012 A1 of the year 2009 called "Protein-based probiotic encapsulates" teaches an encapsulation that has a protein-based encapsulation matrix, which involves one or more preferably Gram-positive probiotic bacteria; These bacteria have an average mass of 1-5000pm, where the encapsulation matrix contains at least 10% w / w of a protein that has been crosslinked with disulfide bonds. W02009 / 070012 A1 agrees with the present invention because it teaches probiotic compositions that include Lactobacillus acidophilus, L casei, L. rhamnosus; Bifidobacterium breve, B. infantis, B. longum and Streptococcus thermophillus, and mentions the possible addition of powdered milk, trehalose, vitamins, gum arabic and acetate. However, W02009 / 070012 A1 does not disclose that the process includes the addition of the Opadry product or the incorporation of vitamin E, specifically. Similarly, the granulation and subsequent coating process that are incorporated in the present invention are not suggested or mentioned herein. The objective of W02009 / 070012 A1 is to provide the gastric resistance of the supplied probiotic, while the present invention seeks storage at room temperature and its shelf stability.

On the other hand, patent document US2011 / 0268703 A1 of the year 2011, called "Probiotic strain and antimicrobial peptide derived therefrom" shows the study of a strain of Enterococcus mundtii with probiotic characteristics. The E. mundtii strain produces an antimicrobial peptide that exhibits antibiotic activity against a wide range of bacteria. Similarly, a process is disclosed to provide for the production of a peptide comprising culturing the strain in a nutrient medium under microaerophilic conditions at a temperature between 10 ° C and 45 ° C until a recovery of an important peptide. The isolated peptide can be used as an antimicrobial agent in a liquid formulation or a gel formulation as a topical treatment and also as an antimicrobial agent in an encapsulation polymer. However, US2011 / 0268703 does not disclose any method of encapsulation or protection of probiotics, nor the use of protective matrices.

Third, patent document US2014349850A1 of the year 2014, called “Coated dehydrated microorganisms with enhanced stabUity and viability” reveals a technology that seeks the protection of dehydrated microorganisms by adding a layer of hygroscopic salt. Dehydrated coated microorganisms are surrounded by a cover that contains at least 25% hygroscopic salts with a pH that allows the stability and viability of the microorganism. It mentions the existence of at least one cover composed of different materials such as fats, fatty acids, emulsifiers, oils, fats, resins, low permeability polymers, hydrocolloids, starches derived from potatoes, corn, wheat, cassava; polyols and cellulose, as well as the mixture of any of these components. Among the hygroscopic salts mentioned are dipotassium phosphate, sodium phosphate, sodium hexametaphosphate, anhydrous sodium acetate, lithium chloride, magnesium chloride, sodium acetate, calcium acetate and sodium formate among others. Such granules ensure a stability of 15-40 ° C for two years. The proposed technology is through lyophilization or spray coating, the absorption of the microorganism suspended in oil on an inert porous transporter. It also mentions the possibility of bed drying. This foregoing, however, does not mention the formation of granules and their subsequent coating as proposed in this invention. Similarly, the use of hygroscopic salts is not used in the present invention to achieve the same effect.

Another relevant background is the patent document WO2012026832A1 of the year 2013, called "Process ofproducing shelfstable probiotic food", which reveals the coating of fat particles, sugars, waxes, with a hyperosmotic suspension of probiotic microorganisms. In this invention they guarantee the stability of probiotics in food products for 12 months at room temperature. They mention that it is done in fluid beds and granulation is promoted. During the fluid bed process they present as a solid phase, a liquid and a soda interact simultaneously in the chamber of the equipment. Although in this previously, equivalent bed drying technologies are incorporated to those used in the present invention, it does not mention the two phases proposed therein, nor does the use of top spray or tangential nozzles for each stage. They only use top spray in their entire process.

In this review, microorganisms that have been prepared in a culture medium and that have been exposed to stress processes that give them resistance are used as binding agents. In our invention the binding agent is a gum together with stabilizing materials such as Trehalose, Vitamin E, Acetate and whole milk powder. These are the ones that are going to coat the microorganisms that are in a powder mixture and not the opposite as in this patent.

The microorganisms of the present invention are not subjected to any dilution or mixing process in liquids, nor exposed to stress conditions. This patent mentions that the loss of the viability of probiotics over time is approximately two logarithmic units after 6 months at room temperature. In the present invention, the total count is assured without losing logarithmic units. Already by microbial genera, Lactobacilli and coconuts remain in the same logarithmic unit, while Bifidobacteria lose two logarithmic units after 12 months.

On the other hand, the document of US20130115334A1 of the year 2013, called "Heat resistant probiotic compositions and healthy food compressing them" reveals the production of multilayer particles of probiotics, in which granules of probiotic microorganisms are generated to be mixed in healthy foods and Resist high temperatures. Foods include cakes, bread, dairy products and others. Describe a compound granule by different layers such as a probiotic core, an inner oily layer and two outer layers composed of polysaccharides. The first layer is of dried microorganism either by wet route, by dry granulation or by hot melt granulation. The second layer that is oily can be generated with waxes, fatty acids, fats or oils to generate a moisture protective layer. The first outer layer is aimed at resistance to gastrointestinal conditions. And the last layer aims at thermal resistance and is achieved through the incorporation of a fibrous or gelatinous polysaccharide. Opadry can be used on the outer cover. Although some starting compounds present in the present invention are disclosed in this document and the use of a fluid bed dryer is mentioned, what is sought is to be able to withstand baking and food preparation temperatures, but does not mention stability in time or much less in shelf storage conditions. The above also does not describe the type of spray used in the different drying processes for the formation of the layers. Similarly, in this patent the formation of the first layer or center of the granule containing the probiotics can be similar in terms of the materials used, but without even suggesting the use of whole milk powder which is the main component or support of the present invention. It is then granulated and in the present invention the binder and stabilizer material is used to make the wet granulation. In this patent they use a granulation technology called hot melt that is based on the coating with a molten oily layer (grease, oil, wax). Subsequently, they generate two layers whose objective is gastrointestinal protection and resistance to high temperatures. On the contrary, in the present invention the coating has the objective of protecting moisture so that it can be stored on a shelf at room temperature.

Another background of interest is the patent document WO2016149636A1 of the year 2016, called "Stable granules with low internal water activity", which reveals a process of granulation and coating of particles by which stable granules are obtained, with a low internal water activity. Such granules are suitable for applications in steam treatment processes, in tablet formation and food processing.

The process in question is focused primarily on the stabilization of enzymes for use in animals, although probiotics are mentioned, and seeks stabilization against pelletizing and food processing at temperatures between 70 ° C and 95 ° C. However, although wet and dry granulation is mentioned in the process disclosed above, no specific drying technology is incorporated as part of said process, nor are operating conditions or stability in the finished product described, as It only shows the maintenance of the enzymatic activity of the granules during the granulation process but not after it.

Finally, the patent document W002002058735A1 of the year 2002, called "Method of preparing biological materials and preparations produced using same" reveals the addition of a suspension of cells and proteins to coat preexisting particles and final immobilization with materials such as coatings. It is mainly focused on the stabilization of proteins and moisture sensitive materials such as living cells. This technology comprises several stages, the first the generation of a water-soluble cover, then the generation of water-soluble gel particles and ends in a fluid bed drying process to remove moisture. It proposes several stages of coating, enteric, covering and repulsion of moisture or flavoring. The process involves the atomization of a liquid that contains the material of interest with a sugary polymer and a water soluble solvent inside an excipient that is fluidized in a chamber at a temperature greater than the ambient one. The proposed process is carried out in a bed dryer with some modifications both in the spray guns, and in the return of the air, generating a continuous process. Similarly, a bottom spray drying is proposed due to the location of the spray guns. The microorganisms tested are only stable at 2 months.

Given the improvements in the equipment used in this invention, adaptations in the machines and in the process are required. The present invention takes advantage of the spray gun alternatives using top and tangential spray without any modification and ensuring stability for periods longer than the two months mentioned in this reference.

Based on the limitations of the state of the art and the need to provide storage conditions for probiotic-based products for prolonged periods and at high temperatures, the present invention reveals a process that integrates probiotic drying and coating technologies, the application of which allows that the organisms thus microencapsulated and the final products that comprise them, can be stored at room temperature for long periods, without losing their viability and stability, which optimizes their logistic management throughout their distribution process and delivery to the final consumer, even allowing its handling in shelves or gondolas without the need for air conditioning or refrigeration.

DESCRIPTION OF THE INVENTION

The present invention describes a method of obtaining microgranules suitable for transporting any type of sensitive microorganism, including but not limited to probiotic microorganisms, in particle sizes between 100 and 800 microns, which includes the stages of drying, granulation and coating, in a fluid bed dryer.

The application of the claimed process provides coated probiotics that are stable and effective at room temperature for long periods of time without affecting their viability, on shelf storage, even at high temperatures without the need for refrigeration or air conditioning, ensuring period stability prolonged time of up to 12 months. In the same way, the present invention generates a protective layer against moisture and oxygen which allows microorganisms to be stored without affecting their viability and performance.

For a better understanding of the claimed subject matter, the following definitions are included: a. Microencapsulation: It is an operation that involves surrounding a solid, liquid or gaseous base (core) with a sufficiently resistant and stable envelope, immiscible but adherent with the core, and that is only altered by releasing its content in certain media. The size of the units varies between 0.5 and 5000 microns.

b. Cover: refers to a layer that surrounds the microorganism or granule and is applied in order to protect the inner layer or dry core, as the case may be.

c. Drying fluid bed: it is a type of particle drying whose basic principle is the atomization of a suspension in a bed of particles fluidized by hot air injection. During fluidization the water evaporates leaving dry particles. Several drying stages can be generated generating coating layers. d. Lyophilization: is a type of particle drying that aims to remove water from a particle by means of freezing and sublimation at reduced pressures. This process is carried out under vacuum and at low temperatures thus avoiding protein denaturation.

and. Aqueous activity: defined as the relationship between the vapor pressure of a food in relation to the vapor pressure of water at the same temperature. Water activity (3w) is the amount of free water in the food, that is, the water available for the growth of microorganisms and so that different chemical reactions can be carried out. It has a maximum value of 1 and a minimum value of 0. The lower this value, the better the product will be preserved. Water activity is related to the texture of food: at a higher activity, the texture is much more juicy and tender; however, the product is altered more easily and more care must be taken. F. Humidity: refers to the total amount of water present in a product, although it may not be free to interact, which makes it different from the water activity.

g. Probiotic: according to the World Health Organization, probiotics are "Living microorganisms that, when supplied in adequate amounts, promote health benefits of the host organism." These microorganisms can be bacteria and yeasts. The best known genera are Lactobacillus sp., Bifidobacterium sp., Streptococcus sp., Bacillus sp., And yeasts of the genus Saccharomyces sp. With the foregoing in mind, it is mentioned that the claimed process is constituted by the following general stages, which are described as an illustration without limiting the scope of the present application: a. Manufacture of a core by wet granulation by fluid bed: A dry mixture of probiotics and powdered milk is granulated by the addition of a binder solution (gum arabic) and stabilizing materials (Trehalose, Vitamin E Acetate and whole milk powder). This granulated nucleus is characterized by containing low levels of aqueous activity and moisture. b. Then, the granulate obtained is subjected to a coating process with a film that allows the core to be isolated from environmental conditions such as humidity and oxygen. This coating is also carried out in a fluid bed by atomizing a coating solution. c. At the end of the process, granules coated with probiotic microorganisms with sizes between 100 and 800 microns, a humidity between 3% and 5% and an aqueous activity between 0.3 and 0.5 can be suspended and can be suspended in water.

BRIEF DESCRIPTION OF THE TABLES.

Table No. 1 Counting and survival of microencapsulated microorganisms

It shows the microbial count expressed in colony forming units cfu / g of acid-lactic bacteria over time, as well as the survival percentage thereof up to 12 months.

Table No. 2 Microencapsulated microorganism count by gender

It shows the microbial count expressed in colony forming units cfu / g of bacteria of the genus Lactobacillus sp., Bifidobacterium sp., And coconuts that were microencapsulated and stored for 12 months.

Table No. 3 Survival of microencapsulated microorganisms by gender

It shows the survival of bacteria of the genus Lactobacillus sp., Bifidobacterium sp., And coconuts that were microencapsulated and stored for 12 months. Expressed as a percentage of survival.

BRIEF DESCRIPTION OF THE FIGURES

FIGURE No. 1. Microencapsulated acid lactic acid count

It shows the microbial count expressed in colony forming units cfu / g of acid-lactic bacteria over time, in a graphical way over time for 12 months.

FIGURE No. 2 Microencapsulated bacteria count by gender

It shows the microbial count expressed in colony forming units cfu / g of bacteria of the genus Lactobacillus sp., Bifidobacterium sp., And coconuts that were microencapsulated and stored for 12 months graphically.

FIGURE No. 3 Survival of microencapsulated probiotics

It shows the survival of microencapsulated and stored acid-lactic bacteria for 12 months. Expressed as a percentage of survival graphically.

FIGURE No. 4 Survival of microencapsulated probiotics by gender

It shows the survival of bacteria of the genus Lactobacillus sp., Bifidobacterium sp., And coconuts that were microencapsulated and stored for 12 months. Expressed as a percentage of survival graphically.

Figure No. 5 Electron microscopy of granules before and after the microencapsulation process

It shows by electron microscopy the appearance of the granules before and after the microencapsulation process in different approaches, A. 4000 X, B. 2100 X, C. 1800 X, D. 1400 X and E. Before microencapsulation 1100 X. EXAMPLE 1.

MIXTURE

In a mixer the total of probiotics and whole milk powder are added, in a 1: 2 ratio (milk: probiotics). The order of addition is given by the density of the materials, start with the load of higher density material and end with the one with the lowest density. Mix between 15 and 40 minutes. The final mixture is called Dry Core.

EXAMPLE 2

PREPARATION OF THE BINDING SOLUTION

In a container with constant agitation, the gum arabic is hydrated for 1 to 6 hours. This is done with water and is brought to a concentration of 25%. The ratio between the rubber and the dry core should be stored at 1: 6. After obtaining the homogeneous dispersion and maintaining constant agitation, the stabilizers are added: trehalose that can be incorporated in proportions between 1: 50, 1: 40, 1: 30 or 1: 20 in relation to the dry core; whole milk powder that can be incorporated in proportions against the dry core between 1: 5, 1: 7 or 1: 9 and vitamin E acetate that can be incorporated between 1: 40, 1: 50, 1: 60 and up to 1: 80 in proportion to the dry core. Stirring is continued for 30 to 60 minutes until all materials are properly incorporated to obtain a lump-free fluid dispersion. The gentle agitation is continued during the process. EXAMPLE 3

GRANULATION

After loading the dry core in the fluid bed dryer, the granulation process occurs, consisting of the top spray spray of the binder solution on the probiotic mixture.

The granulation process is preferably carried out in a fluid bed equipment with top spray atomization gun arrangement. The core is controlled to grow to an approximate average size between 100 and 800 microns. A temperature in the fluidization chamber is handled between 30 ° C and 40 ° and a total process time of approximately 300 minutes.

This process is composed of a stage of heating and fluidization of the dry core, followed by the atomization of the binder solution. After the total addition of binder, it is dried for 30-60 minutes or until a humidity less than or equal to 5% and an aqueous activity less than or equal to 0.5 is reached. The fluidization of materials is carried out with air flow between 700 and 3000 m ³ / h and an inlet air temperature not exceeding 45 ° C.

It must be ensured that the product temperature during the entire atomization does not exceed 35 ° C. If a decrease in the fluidization of the product is observed, increase the flow of inlet air or minimize the addition of the binder solution and let it dry for a few minutes until the proper fluidization of the product is obtained again. EXAMPLE 4

COVERING

In a container provided with stirring, sufficient purified water is added to achieve a 23% (w / w) aqueous dispersion of Opadry 200 under permanent agitation for 30 to 60 minutes. The coating process is carried out in a multifunctional fluid bed system with a spray tangential spray gun arrangement. It is sought to cover the granule in a controlled manner until it reaches its total coverage. A temperature is handled in the fluidization chamber that does not exceed 40 ° C, with inlet air not exceeding 60 ° C and with an air flow between 1600-3000 m ³ / h. The total process takes approximately 300 minutes.

After the addition of the coating film, it is dried for 30-60 minutes or until a humidity less than or equal to 5% and an aqueous activity less than or equal to 0.5 is reached. The granule maintains its size generated in the granulation, that is 100 to 800 microns. At the end of this stage, a sample should be taken and the moisture content and the water activity must be verified, which must be a maximum of 5% and a 3w of maximum 0.5 respectively.

After this stage the equipment is unloaded and the container of the microencapsulated material.

Table No. 1 Counting and survival of microencapsulated microorganisms

Table No. 2 Microencapsulated microorganism count by gender

Table no. 3 Survival of microencapsulated microorganisms by gender

Claims

1. A process for coating and encapsulating probiotics and other materials sensitive to moisture and oxygen, characterized by comprising the steps of:

(i.) Manufacture a core by wet granulation by fluid bed.

(ii.) Coat the granulated core by atomizing a coating solution.

2. The method of claim 1, characterized in that the manufacturing of the granulated core comprises the steps of:

(i.) Dry mix, for a period between 15 and 40 minutes, the selected probiotic microorganisms with whole milk powder, in a ratio of 1: 2 (milk: probiotics).

(ii.) Prepare, by agitation for times between 1 and 6 hours, a binder solution formed by gum arabic dissolved completely in water and brought to a concentration of 25%, where the proportion between the rubber and the dry core is 1: 6, obtaining a homogeneous dispersion;

(iii.) Maintain constant agitation on the dispersion and add trehalose in proportions between 1:50, 1: 40, 1: 30 or 1: 20 in relation to the dry core;

(iv.) Incorporate whole milk powder in proportions of 1: 5, 1: 7 or 1: 9 in relation to the dry core, and vitamin E acetate in proportions of 1: 40, 1: 50, 1: 60 and 1 : 80 in relation to the dry core. (v.) Maintain stirring for 30 to 60 minutes until the incorporation of all materials ensures a smooth lump free dispersion.

(vi.) Granulate the microorganisms by atomizing the binder solution by top spray spray until obtaining a dry granulated nucleus of the selected microorganisms; (vii.) Load the dry granulated core, atomize the binder and fluidize the entire mixture in the dryer at a maximum temperature of 35 ° C for a maximum total time of 300 minutes, with air flows between 700 and 3000 m3 / h input air to the equipment at a maximum temperature of 45 ° C.

3. The method of claim 1, characterized in that the coating of the granulated core by atomization comprises the steps of:

(i.) Prepare a film-forming solution, consisting of polyvinyl alcohol in water at a concentration of 23% and keeping a proportion in front of the dry granulated core of 1: 3 (coating agent: granulated core)

(ii.) Coat the granule by atomizing the solution on the granulated core, by drying a fluidized bed with tangential atomization by atomizing the coating film on the granulated core and fluidizing this mixture, applying atomization pressure of 0.3 bar, with inlet air at a maximum temperature of 50 ° C and an air flow between 1600 and 3000 m ³ / h, without allowing coated granules to reach temperatures above 35 ° C.

4. The granulated and coated probiotic obtained according to the procedure of the preceding claims, characterized in that it has a size between 100 and 800 microns, with a maximum humidity of 5% and an aqueous activity not exceeding aw 0.5.

5. The granulated and coated probiotic of the preceding claim, characterized in that it is capable of being suspended in water or other liquids at temperatures between 20 ° C and 35 ° C.

6. The granulated and coated probiotic of the preceding claim, characterized by ensuring survivals greater than 70% for 12 months when stored at room temperature of up to 30 ° C.

7. The granulated and coated probiotic of claims 4 to 6, characterized in that it is suitable for inclusion in different food and pharmaceutical matrices, including but not limited to soluble powders, sachets, capsules and tablets or other solid pharmaceutical forms and / or powders Solid nutritional