TECHNICAL FIELD

The invention relates to an encapsulation that enables probiotic microorganisms to form a high level of colony in the gastrointestinal tract in the human body by becoming more resistant to external conditions and to make controlled release, to create probiotic food products by remaining alive in various food processing processes, and a method for the encapsulation of the said probiotic microorganisms and the use of encapsulations comprising the said probiotic microorganism in food products.

BACKGROUND

Probiotic microorganisms are beneficial bacteria and yeasts that help various body systems, especially the digestive system and skin, to work in a balanced manner, unlike bacteria that cause infection and diseases. The beneficial bacteria, which also play a great role in eliminating the disease-causing effects of harmful bacteria, are naturally present in the body. The human body has a system called the microbiota/microbiome outside the cells. All microbes in the human body (bacteria, fungi, viruses, protease, etc.) are part of this system, and each individual's body microbiome is different. The beneficial bacteria in the microbiome are microorganisms that contribute to health in the human body, continue to live in internal organs and do not harm to be consumed. Due to these properties, probiotic microorganisms, whose numbers have decreased in the human body for any reason, should be able to be supported with various supplements.

In order to achieve the desired effects from probiotic microorganisms, they should be colonized in sufficient amounts in the gastrointestinal tract (to be abbreviated as GIS). However, probiotic microorganisms consumed orally with food and/or as supplementary food cannot form a sufficient level of colony in humans due to human GIS conditions. Therefore, these effects of probiotic microorganisms cannot be achieved at the desired level. There are four pathways that probiotics taken into the body in the form of supplementary food or functional food potentially follow; (1) some of the oral probiotics are broken down due to low pH conditions of the stomach, (2) those that pass into the small intestine by getting rid of acidic conditions of the stomach lose their vitality due to bile salt, (3) some are destroyed by the normal flora of the small intestine, and (4) those that cannot be destroyed can be recovered from the stool by colonization. If a probiotic microorganism is recovered from stool in amounts close to those consumed, that probiotic microorganism will be able to colonize humans in the required proportions.

On the other hand, another probiotic consumption source of consumers is functional foods comprising probiotics. However, nearly 90% of the probiotic foods on the market are in the form of milk and dairy products. The biggest reason for this is the structure of probiotic microorganisms that are not resistant to external conditions and this structure does not allow the formation of functional foods comprising various probiotics.

The presence of acrylamide in food and beverages was announced by the SNFA (Swedish National Food Commission) in 2002 and has been included in the 2A carcinogenic (highly carcinogenic) substance group for humans by the IARC (International Institute for Research on Cancer). Acrylamide is formed when sugars such as glucose and fructose in green coffee beans are reduced by the high temperature effect they are exposed to in the roasting process and react with Asparagine. OTA, on the other hand, is a toxic metabolite produced by Penicillium Verrucosum and Aspergillus Ochraceus microorganisms and has been identified by IARC as a 2D carcinogen (probably carcinogenic in humans) for humans. In various studies, it has been revealed that the sources where people are most exposed to Acrylamide and OTA during the day are coffee types, while coffee consumed by a person worldwide on average constitutes 25% of the daily Acrylamide intake and 7% of the daily intake of OTA. However, it has been reported in various studies that the amount of acrylamide that people can tolerate daily is 0.3-0.6 pg/kg and the amount of OTA is 1 .5-5 ng/kg. However, acrylamide and OTA intake sources are not limited to coffee. While sources such as red peppers, biscuits, chocolate, alcohol products and rice are intermediaries for the purchase of HERBS, various heat-treated junk foods, bread, pastry products, french fries, red meat and potato chips are intermediary sources for the purchase of acrylamide.

As a result, it is necessary to make innovations in order to increase the effectiveness of probiotic microorganisms known to have technical benefits for the human body in the relevant technical field to form a high level colony in the human body without being affected by GIS conditions, to qualify various foods with probiotics by providing maximum probiotic viability in the food processing process, and to eliminate the substances called acrylamide and Ochratoxin A in the food with probiotic microorganisms. BRIEF DESCRIPTION OF THE INVENTION

The object of the present invention is to enable the probiotic microorganisms to become more resistant to outdoor conditions.

The object of the present invention is in the hands of probiotic microorganisms that exhibit high resistance under human body GIS environmental conditions.

The object of the present invention is to provide controlled release of probiotic microorganisms under human body GIS environment conditions.

The technical problem determined for the relevant technical field is that the probiotic microorganisms, whose technical benefits are known, do not have sufficient strength to form a colony at the desired levels under GIS environment conditions and cannot be placed in the GIS environment with controlled release. The present disclosure relates to the disclosure of nano- and/or micro-sized encapsulations for probiotic microorganisms that will provide resistance to GIS environmental conditions against all these technical drawbacks. Furthermore, the present invention relates to a method for encapsulating probiotic microorganisms with technical benefits with micro and/or nanoscale encapsulations that will provide durability.

In order to provide all these, the invention relates to an encapsulation comprising components of chocolate, starch, inulin, maltodextrin and enabling probiotic microorganisms to form a high level of colony in the gastrointestinal tract of the human body by being made more resistant to external conditions and to produce probiotic food products by remaining alive in various food processing processes.

The possible embodiment of the invention is that the encapsulation further comprises a cellulose acetate phthalate component in it.

In a preferred application, the encapsulation comprises a chocolate component in the range of 30-70% by weight. In another preferred application, the encapsulation comprises within itself a chocolate component in the range of 40-50% by weight of the chocolate component. In the most preferred application, the encapsulation comprises a chocolate component in the range of 45 to 49% by weight. In a preferred application, the encapsulation comprises a starch component in the range of 1% to 10% by weight in the body.

In a preferred application, the encapsulation comprises a maltodextrin component in the range of 1% to 10% by weight.

In a preferred application, the encapsulation comprises an inulin component in the range of 1% to 10% by weight.

In a preferred application, the encapsulation comprises a cellulose acetate phthalate component in the range of 0.5% to 5% by weight.

In a preferred application, the encapsulation comprises a starch component in the range of 1% to 10% by weight, a chocolate solution component in the range of 30% to 70%, a maltodextrin component in the range of 1% to 10% by weight, an inulin component in the range of 1% to 10% by weight, a maltodextrin component in the range of 1% to 10% by weight, and a cellulose acetate phthalate component in the range of 0.5% to 5% by weight.

The invention also relates to a method for the production of probiotic microorganisms, encapsulation. The inventive encapsulation method includes the following steps: i) Applying the emulsification process step and obtaining the pre-encapsulation

A mixture of microorganism-coating materials is obtained by homogenizing the coating materials sterilized with multiplied probiotic microorganisms, sunflower oil is added during this process,

The mixture of homogenized microorganism-coating materials is dropped into a stabilization solution with the help of a syringe and the mixing process is carried out by means of a mixer,

The CaCh solution is slowly added and the mixing process is continued to increase the stability of the mixture,

The filtration of the solution and pre-encapsulation are obtained as a result of mixing processes, ii) Chocolate encapsulation and freeze-drying

The anterior encapsulations are completed to at least 1 liter with water and then filtered, The obtained filtrate is kept in the range of 10 seconds to 120 seconds at a temperature range of 60 to 75°C by the heater and then cooling operations are carried out using ice,

The chocolate solution is prepared in a separate portion, wherein the said chocolate solution is added to water and/or PBS solution and mixing operations are carried out at a temperature between 80 and 95°C, then the said chocolate-water and/or PBS solution is allowed to cool,

The pre-encapsulations obtained as a result of the processes are added to the chocolate solution and a homogeneous mixture is obtained by mixing at least 100 rpm for at least 30 minutes,

The filtered products are subjected to freeze-drying to obtain encapsulated products.

In a possible embodiment of the invention, the ratio of the materials used in the preencapsulation process in the process step i) has a value in the range of 1 :5 to 1 :10 (ml:ml) as microorganisrmpre-encapsulation materials.

In a possible embodiment of the invention, the chocolate solution preparation process in the process step ii) includes water and PBS components together.

In the possible embodiment of the invention, the pH of the said solution is preferably in the range of 7 to 7.5.

In a possible embodiment of the invention, in the process step ii), the chocolate solution is added such that i) the pre-encapsulation/chocolate solution is in the range of 1/5 to 1/15 (g/ml) according to the pre-encapsulations obtained in the process step i).

In the possible embodiment of the invention, the cryopectane sorbitol compound is added in the process of freezing and drying the process step ii).

In the possible embodiment of the invention, 1% by weight cryopectane sorbitol compound is added in the process of freezing and drying the process step ii).

DETAILED DESCRIPTION OF THE INVENTION

The subject of the invention is a method for the encapsulation of probiotic food products by making the probiotic microorganisms more resistant to external conditions and allowing the probiotic microorganisms to form a high level colony in the gastrointestinal tract and to make controlled release in the human body, and a method for the encapsulation of the said probiotic microorganisms by remaining alive in various food processing processes and the use of encapsulations comprising the said probiotic microorganism in food products and is explained with examples that do not have any limiting effect only for a better understanding of the subject.

In the invention, the term "probiotic microorganism" refers to bacteria or fungi that help the human body's various systems, especially the digestive system and the skin, work in a balanced manner. The subject of the invention is independent of the type of probiotic microorganism included in the encapsulation and encapsulation method of the invention. Preferably, beneficial bacteria for the human body are used as probiotic microorganisms. Examples of beneficial bacteria include bacteria from the Lactobacillus, Streptococcus and Bifidobacterium groups.

In the invention, "gastrointestinal system" refers to a long tubular structure between the mouth and the anus and a system that includes many organs associated with this structure and whose main task is digestion. It is expected that the probiotic microorganism encapsulations of the invention will preferably remain intact in the gastrointestinal tract and that the probiotic microorganisms contained therein will be released after a while.

In the invention, "encapsulation" is the technology that protects a very small substance or components by coating solid, liquid, and gas materials for capsules and allows them to be released under certain conditions and at certain speeds. In the invention, probiotic microorganisms known to have technical benefits are encapsulated in order to be resistant to GIS environmental conditions in the human body and process environmental conditions in order to be used in foods and to allow controlled release in the human body after a while.

The encapsulations of the invention are preferably added to food products and thus it is possible to obtain food products comprising probiotic microorganisms. Encapsulations comprising probiotic microorganisms to be added to food products should also have sufficient resistance to outdoor conditions such as high temperature or mechanical forces to be realized during the production of food products. The encapsulation of the invention comprises coating materials to ensure that the probiotic microorganisms remain alive as much as possible in the food and under GIS environment conditions. Accordingly, the encapsulation comprises components that adapt to GIS external environment conditions both during food technology production and after incorporation. The encapsulation of the present invention comprises at least one, several or all of soybean lestine, inulin, chocolate, whey protein, starch, maltodextrin, cellulose acetate phthalate substances.

The encapsulation of the invention preferably comprises chocolate, high amylase starch, inulin, maltodextrin, cellulose acetate components all at once.

The encapsulation of the invention comprises a chocolate component in the range of 30- 70% by weight. The said chocolate is preferably dark chocolate. In a preferred application, the encapsulation comprises a chocolate component in the range of 40-50% by weight. In the most preferred application, the encapsulation comprises a chocolate component in the range of 45 to 49% by weight. The chocolate component contained in the encapsulation of the invention allows the protection of probiotic microorganisms and resistance to heat treatment. If the chocolate component has sub-values of 30% by weight in the invention, it will be insufficient for the formation of the encapsulated structure. In the invention, if it comprises more than 70% by weight of chocolate components, it causes the encapsulations to have very large structures. In order to obtain the encapsulation suitable for the desired strength values and ambient conditions, it is essential that the chocolate component is in the determined weight ratios. The chocolate component is included in the encapsulation and thanks to its sweetening, it can be easily added to food products. In addition, the chocolate component can also be included in a food product that people can easily consume thanks to its encapsulation.

The encapsulation of the invention comprises a starch component with a high amylase content in the range of 1% to 10% by weight. The said starch may have been manufactured from wheat, corn or barley. In a preferred application, the encapsulation comprises starch made from corn. It is essential that the amylase-containing starch component is in the proportions by weight in order to obtain the encapsulations suitable for the desired strength values and the ambient conditions.

The encapsulation of the invention comprises an inulin component in the range of 1% to 10% by weight. In order to obtain the encapsulation suitable for the desired strength values and ambient conditions, the inulin component should be in the said weight proportions.

The encapsulation of the invention comprises the maltodextrin component in the range of 1 to 10% by weight. The encapsulation of the invention comprises a cellulose acetate phthalate component in the range of 0.5% to 5% by weight, if preferred. This range is preferred in order to increase the efficiency of the encapsulation and to have the desired size.

The invention further relates to a method of encapsulation comprising the components given in the invention and having the configurations for the probiotic microorganisms. The said method includes the steps of i) obtaining pre-encapsulations by emulsification and ii) encapsulating and freeze-drying with chocolate material. i) Applying the emulsification process step and obtaining the pre-encapsulation

Probiotic microorganisms that are previously determined and incubated with known methods in the art are reproduced,

The encapsulation materials are sterilized except for the chocolate component,

A mixture of microorganism-coating materials is obtained by homogenizing the coating materials sterilized with multiplied probiotic microorganisms, sunflower oil is added during this process,

The mixture of homogenized microorganism-coating materials is dripped into a stabilization solution with the help of a syringe and mixed with a mixer,

Slowly adding CaCb solution to increase the stability of the mixture and continuing the mixing process,

Filtering the solution as a result of mixing processes and obtaining the preencapsulations. ii) Chocolate encapsulation and freeze-drying

The anterior encapsulations are completed to at least 1 liter with water and then filtered,

The obtained filtrate is kept in the range of 10 seconds to 120 seconds at a temperature range of 60 to 75°C by the heater and then cooling operations are carried out using ice,

The chocolate solution is prepared in a separate portion, wherein the said chocolate solution is added to water and/or PBS solution and mixing operations are carried out at a temperature between 80 and 95°C, then the said chocolate-water and/or PBS solution is allowed to cool, The pre-encapsulations obtained as a result of the processes are added to the chocolate solution and a homogeneous mixture is obtained by mixing at least 100 rpm for at least 30 minutes,

The homogeneous mixture is filtered,

The filtered products are subjected to freeze-drying to obtain encapsulated products.

The ratio of the materials used in the pre-encapsulation process in the process step i) is 1 :5 to 1 :10 (ml: ml), with the microorganism/pre-encapsulation materials. In order to be more descriptive, the value expressed here may be in the range of 5 ml to 10 ml of the preencapsulation materials in case the microorganism value is 1 ml.

Pre-encapsulation materials mentioned in the step i) preferably comprise high amylase starch, inulin, maltodextrin and cellulose acetate phthalate components.

In the i) process step, the inulin component preferably comprises a high amylase content starch in the pre-encapsulation in the range of from 1 to 10% by weight, an inulin component in the range of from 1 to 10% by weight, a maltodextrin component in the range of from 1 to 10% by weight.

The process step i) comprises a cellulose acetate phthalate component in the preencapsulation body, preferably in the range of 0.5% to 5% by weight.

The sterilization processes in the process step i) are carried out at a value in the range of temperatures from 100 to 150°C.

The sunflower oil mentioned in the process step i) is used as a mobile phase component in the system. For encapsulations, the sunflower oil is used partially for stabilizing the surface tension and trapping it in the system.

It is preferred that the inner diameter of the syringe used in the process step of adding to the stabilization solution with the help of the syringe mentioned in the process step i) is preferably 100 mm. Preferably, the pre-encapsulation material with the syringe from a height of 0.7 meters will be dripped into the stabilization solution prepared in the magnetic stirrer, while the stirring process is applied at +4°C and a value in the range of 500 to 1500 rpm by means of the magnetic stirrer during dripping. The chocolate solution preparation process in the process step ii) includes the water and the PBS components together. Preferably, the pH value of the obtained water-PBS- chocolate solution is in the range of 7 to 7.5.

In the process step ii), the chocolate solution is added so that the pre- encapsulation/chocolate solution is in the range of 1/5 to 1/15 (g/ml) according to the preencapsulations obtained in the process step ii). In order to be more descriptive, the value expressed here may be in the range of 50 ml to 150 ml of the chocolate solution used if the pre-encapsulation value is 10 g.

In the invention, in the process step ii) in the freeze-drying process, 1% by weight of cryopectane sorbitol will be added. The object of the addition of cryopectane sorbitol is to ensure that the microencapsulations formed do not deteriorate during the freeze-drying phase, do not open and do not lose their cell viability due to osmotic pressure.

The encapsulation obtained by the application of process steps i) and ii) has a particle diameter of 1 to 500 micrometers. In this respect, the encapsulation with a very low particle size diameter can be obtained in the present invention.

With the components contained in the encapsulation of the invention and the method of encapsulation of probiotic microorganisms, probiotic microorganisms can be eliminated with probiotic microorganisms by both making them resistant to external conditions and providing controlled release ability; i) forming a high level of colony in the human body without being affected by the gastrointestinal system conditions ii) providing maximum probiotic viability in the food processing process and qualification of various foods with probiotics iii) disposing the substances called acrylamide and ochratoxin A in the food can be achieved with probiotic microorganisms.

The scope of protection of the invention is specified in the attached claims and cannot be limited to those explained for sampling purposes in this detailed description. It is evident that a person skilled in the art may exhibit similar embodiments in light of above-mentioned facts without drifting apart from the main theme of the invention.