BRPI0905590A2 - industrial process of immobilization of a consortium of microorganisms present in kefir biologicus, as well as their bioactives, through the formation of modified calcium alginate microcapsules - Google Patents

Abstract

INDUSTRIAL PROCESS OF IMMOBILIZATION OF A CONSORTIUM OF MICRO-ORGANISMS PRESENT IN KEFIR BIOLOGICUS, AS WELL AS ITS BIOACTIVES, THROUGH THE FORMATION OF MODIFIED CALCIUM ALGINATE MICROCASULES where microorganisms are present in their conservation, as well as microorganisms present in the kefir, thus without the care required by unprocessed kefir, as well as enabling its inclusion in foods, cosmetics and medicines without changing their color, taste or odor and, at the same time, giving them the probiotic benefits of kefir.

Description

"INDUSTRIAL PROCESS OF! MOBILIZING A MICRORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS AS WELL AS ITS BIOACTIVES BY MEASURING CALCIUM MODIFIED ALGINATE MICROCapsules".

The present invention deals with an industrial process of

immobilization of a consortium of microorganisms present in KefirBioLogicus, as well as their bioactives, by means of spherical microcapsule encapsulation based on Kefirane and Pectin modified calcium alginate. Such invention allows the distribution of microorganisms and their bioactives without the need for greater maintenance by the user, as well as the direct application of microcapsules in food, cosmetics and dermocosmetics, making them probiotic and / or with probiotic components.

In the early 1970s, Todd defined microencapsulation as packaging technology, with fine polymeric coatings applicable to solids, deliquid droplets, or gaseous material, forming small particles called microcapsules, which can release their contents at specific speeds and conditions. In turn, ARSHADY (1993) described microcapsules as extremely small packages, composed of a polymer as a wall material and an active material called a core. Also according to SHHIDI and HAN (1993), microcapsules have the ability to demodify and improve the appearance and properties of a substance. These authors listed the following reasons for the use of microencapsulation in the food industry: (a) reducing the reactivity of the core material to the environment; (b) decrease the rate of evaporation or transfer of encapsulated material; (c) promote controlled release; (d) mask unpleasant odor flavors; and (e) promote homogeneous dilution of encapsulated material in a food formulation. In recent years, such definitions and uses of microencapsulation have been expanded due to the new needs that the food industry demonstrates in increasingly complex properties in formulations, which can often only be met by such a process (GOUIN, 2004). Within these needs, the concept of controlled release has been increasingly requested (GOUIN, 2004; SANGUANSRI and AUGUSTIN, 2006; UBBINK and KRUGER, 2006). There is a worldwide trend that points to the need for food to no longer be seen as a source of nutrients with sensory appeal, but also as a source of well-being and health for the population. This change of perspective undoubtedly requires paradigm shifts in development of new products, applying the traditional methods, but also observing the need to control the bioaccessibility of certain food components. This approach becomes increasingly relevant as the relations between delivery, food and health are established (SANGUANSRI and AUGUSTIN, 2006), and microencapsulation is an effective means of achieving such goals. Active ingredients added to foods include traditional ingredients such as aromas, vitamins, minerals, and also newer ones such as probiotic microorganisms (FAVARO-TRINDADE and GROSSO, 2002; KAILASAPATHY, 2006), bioactive peptides (ARIMOTO ET AL., 2004) and several other classes of bioactive. GOUIN (2004) and DESAI and PARK (2005) have even stated that, through finely tuned controlled release properties, microencapsulation ceases to be merely a method of aggregating substances into a food formulation, and becomes a source of totally new ingredients as unique properties. . In addition, innovation in the food industry already undoubtedly already needs a shift in focus from observing macroscopic properties to meso and nanoscale eateries, with subsequent control of hierarchical structures in foods and their functionality. For this reason, studies on the relationship between nano and supramolecular structures are increasingly essential, as well as functionality at the physical, nutritional and physiological levels (SANGUANSRI and AUGUSTIN, 2006).

Probiotic microorganisms are, in the World Health Organization's concept, "living organisms which, when administered in adequate amounts, confer a benefit on the health of the host." Probiotics, for other authors, are defined as microorganisms that settle in the gut and provide beneficial effects to the host (human or animal), substantially through the maintenance and improved microbiota (between beneficial and harmful microorganisms) of the intestine (FULLER 1989; 1991 GOLDIN 1998, GISMONDO ETAL 1999). Mankind has been using hundreds of years of probiotic foods, especially those derived from milk, such as yogurt, kefir, among others. In recent years, with increasing public awareness of the health benefits of probiotics, there has been market demand for this type of food, predominating in the market for milk fermented by one or two types of microorganisms. Various health benefits have been attributed to probiotics as well as antimutagenic, anticancer, anti-inflammatory properties, immune system stimulation, cholesterol reduction, relief of lactose and lactose intolerance (GILLILAND and SPECK, 1977; KIM and GILLILAND, 1983; RASIC and KURMANN, 1983). ; GURR1 1987; GILLILAND, 1989; SURAWiCS ET AL., 1989; FULLER, 1992; BUCK and GiLLiLAND11994; LANKAPUTHRA AND SHAH1 1995; DALY AND DAVIS1 1998; KLEIN ET AL., 1998; MACFARLANE and CUMMINGS, 1999; 2000). Lactobacillusacidophilus, L. casei, Bifidobacterium bifidum, B. longum, B. brev, B. B. Iactis (B. animalis) are the most popular bacteria applied to probiotic food products (DALY and DAVIS, 1998; KLEIN ET AL., 1998; MACFARLANE and CUMMINGS, 1999). Much research involving probiotic viability and survival studies in gastrointestinal tract and food products (especially in dairy products) has revealed that viability generally decreases dramatically due to exposure to harmful environmental factors as well as organic acids, hydrogen ions, molecular oxygen and antibiotic components (GILLILAND and SPECK, 1977; HAMILTON-MILLER 1999 IWANA ET AL., 1993; LANKAPUTHRAe SHAH, 1995; SHAH ETAL., 1995; DAVE and SHAH, 1996; DAVE eSHAH, 1997; KAILASAPATHY and RYBKA, 1997; SHAH and LANKAPUTHRA, 1997; KEBARY ET AL., 1998; BEAL ET AL., 1999; GARDINI ET AL., 1999; HAMILTON-MILLER ET AL., 1999; SCHILLINGER, 1999; VINDEROLA ET AL., 2000; SULTANA ETAL., 2000; MORTAZAVIAN ET AL., 2006). In addition, the beneficial effects of probiotic microorganisms appear when they, after surviving the unfavorable conditions, reach the intestines viable and high enough to act in the environment (GILLILAND, 1989). A minimum amount of probiotic cells (CFU / g) in the product at the time of consumption, which is necessary for the beneficial (preventive or therapeutic) pharmaceutical effects of probiotics to be represented by the minimum bio-vaue (MBV) index. -value) (MORTAZAVIAN ET AL., 2006). According to the "International Dairy Federation (IDF)" recommendation this index should be equal to above 107 CFU / g (OUWEHAND and SALMINEN, 1998). In some countries such as Argentina and Brazil this value should be equal to above 106 CFU / g for bifidobacteria. This pattern in Japan has been shown to be above 107 CFU / g (ROBINSON, 1987). Other recommendations have been presented by different researchers, as well as an index above 106UFC / g for all probiotics present in yogurts (ROBINSON, 1987; KURMAN and RASIC, 1991) and above 107UFC / g in the case of bifidobacteria (HOLCOMB ET AL. , 1991). In addition to the MBV index, there is a second index called the "intakedaily" index of each food product also determinable by its probiotic potential or effectiveness. The minimum amount of this index has been recommended with approximately 109 viable cells per day (SHAH ET AL., 1995; KURMAN and RASIC, 1991; MORTAZAVIAN, 2006). Loss of viability of probiotic food products (especially fermented types) and acid conditions of the bile of the gastrointestinal tract have encouraged researchers to find new efficient methods that maintain viability. Microencapsulation, as one of the newest and most efficient methods, has been the subject of special consideration6 / 36

and research. From a microbiological point of view, microencapsulation can be defined as a process of trapping microorganisms by coating them with appropriate hydrocolloid (s) in order to separate them from the medium. This process results in an appropriate release of microorganisms in the intestinal environment (SULTANA ET AL., 2000; KRASAEKOOPT ET AL., 2003; PICOT and LACROiX1 2003a). Among the agents that cause the release are: pH change, mechanical stress, heating, enzymatic activities, pressure osmotic, presence of some chemical components and storage time can be mentioned (GOUIN, 2004). Micro-encapsulation of microorganisms preserves them from the action of medium factors as well as high acidity and low pH (WENRONG and GRIFFITHS, 2000), bile salts (LEE and HEO, 2000), thermal shocks induced by process conditions as well as freezing and cold drying ( SHAH & RARULA, 2000), molecular oxygen in the case of anaerobic microorganisms (SUNOHARA ET AL., 1995), hot thermal shocks caused by process conditions as well as spray drying (STEENSON ET AL., 1987) and antimicrobial chemicals (SULTANA , 2000). Other advantages pointed out are the increased stability of sensory properties and / or their improvement (GOMES and MALCATA, 1999) and the immobilization of microorganisms for their homogeneous distribution in a product (STEENSON ETAL., 1987; KRASAEKOOPT ET AL., 2003). The importance of the microencapsulation method as an efficient way to increase probiotic viability justifies the importance of this application. The microcapsules have characteristic structures. Each microcapsule consists of a hydrocolloid coating (also called a capsule) surrounded by the microorganism (s). Because the microcapsules are spherical or elliptical in shape, they are also called microspheres. The microcapsules may have a smooth or rough surface. Each microcapsule can encapsulate one or more types of microorganisms. When multiple microorganisms are present in the microcapsule, the solution's interstitial fluid fills the voids in the microcapsule. Superficial and / or deep cracks may appear in the microcapsules. The propagation of these cracks leads to pore formation, which considerably reduces the microencapsulation efficiency. The microcapsules may be coated with a second layer of a chemical compound to increase the efficiency of the microencapsulation process. The second layer is also called a coating or support or shield. Microcapsules with or without the presence of a second protective layer are known as coated microcapsules or uncoated microcapsules, respectively. Constituents trapped within the coating are known as the core (SULTANA ET AL., 2000; TRUELSTRUP-HANSEN ET AL., 2002; DIMANTOV ET AL., 2003; KRASAEKOOPT ET AL., 2003; CHANDRAMOULI ETAL., 2004).

There are several components used for the microencapsulation process of probiotics, among them Alginate and its combinations. Alginate is a heteropolysaccharide extracted from different types of algae, with structural units consisting of D-manuronic and L-guluronic acids. Calcium alginate has been widely used for the microencapsulation of lactic acid and probiotic bacteria, mainly in the 0.5-4% concentration range (SHEU and MARSHALL, 1991; SHEU and MARSHALL, 1993; TRUELSTRUP-HANSEN ET AL., 2002; KIM ETAL. 1996; JANKOWSKI ETAL., 1997; KHALIL and MANSOUR, 1998; KEBARY ETAL., 1998; LEE and HEO, 2000; SHAH and RARULA, 2000; SULTANA ET AL., 2000; TRUELSTRUP-HANSEN, 2002; KRASAEKOOPT ET AL. , 2004). Aiginate as an ingredient for microcapsule formation has some advantages as follows (KLIEN ET AL., 1983; TANAKA ET AL., 1984; MARTINSENET AL., 1989; PREVOST and DIVIES, 1992; DIMANTOV ET AL., 2003; CHANDRAMOULI ET AL. , 2004; GOUIN1 2004): easily forms gel around the microorganism, not poisonous to the human body (safe or biocompatible), relatively inexpensive, mild process conditions (as well as temperature) necessary for its performance, can be easily prepared (simplicity and ease in handling) and decomposes properly in the gut and releases the microencapsulated microorganisms. The gelatin gel matrix appropriately surrounds the bacterial cells with a diameter of 1-3μm and the pore sizes formed on the surface of the gelatin microcapsules do not exceed 7nm (KLIEN ETAL., 1983). However, some disadvantages are attributed to the gelatin microcapsules. For example, they are susceptible to acidic environments and their decomposition and loss of mechanical stability in lactic acid containing media have been verified (EIKMEIER and REHM, 1987; ROY ETAL., 1987; AUDET ETAL., 1988; ELLENTON, 1998). When aniginate gel is formed in the presence of calcium ions, its integrity is deteriorated when subjected to monovalent ions or chelating agents, which absorb calcium ions such as phosphates, lactates and citrates (ROY ET AL., 1987; SMIDSROD and SKJAK-BRAEK, 1990; ELLENTON, 1998). Other disadvantages include difficulties in industrial scale application due to its high cost and fragile ability to stagger the control of porosity on the microsphere surface (GOUIN, 2004). This last point leads to a rapid diffusion of the mixture and other fluids through the microcapsules thus reducing its barrier against unfavorable environmental factors (GOUIN, 2004). The effects mentioned can be efficiently compensated for by the use of alginate mixtures with other polymeric compounds, with microcapsule coating compounds and structural modification of alginate by the use of various additives (KRASAEKOOPT ET AL., 2003). Mixing alginate with starch is a common practice and there are some studies showing that the microencapsulation effectiveness of different bacteria, especially lactic acid, can be improved by applying this method (JANKOWSKI ET AL., 1997; SULTANA ET AL., 2000; SUN and GRIFFITHS, 2000 TRUELSTRUP-HANSEN ET AL., 2002; KRASAEKOOPT ET AL., 2003). In addition to good bacterial cell protection, the alginate-starch mixture has the advantage of micronutrient and metabolite diffusion through microcapsules into and out of immobilized organism. As a result, micro particles may contain metabolically active microorganisms (JANKOWSKI ET AL., 1997). Glycerol Alginate increases the survival of microorganisms when stored at very low temperatures, around -20 ° C. This is attributed to the cryogenic effect of glycerol (TRUELSTRUP-HANSEN et al., 2002). The use of a layer over the dreaded alginate microcapsules used to enhance its physicochemical characteristics. It has been reported that a coating with a semipermeable layer of chitosan (a cationic polymer) on the alginate microcapsule (an anionic polymer) increases the physicochemical stability of the produced microcapsules. This structure showed tolerance against deteriorating effects of calcium-burning agents and anti-gelling agents. Also from a structural point of view the microcapsules are denser and more mechanically resistant, thus avoiding the unwanted breaking and release of microorganisms (SMIDSROD and SKJAK-BRAEK1 1990; ZHOU et al., 1998; KRASAEKOOPT et al., 2003). The low molecular weight of chitosan compared to the high pesomolecular alginate causes chitosan to diffuse rapidly through the alginate matrix resulting in the formation of higher density and mechanical strength microcapsules. Calcium chloride coating on dealginate microcapsules has also been investigated (CHANDRAMOULI et al., 2004). The presence of calcium ions generates more stable microcapsules with a greater protective effect on microorganisms and, consequently, greater viability of them. Polyamino acids, as well as Poly-L-Lysine (PLL), are other cationic polymers used as a coating of alginate microcapsules. Similar to chitosan, these polymers form strong complexes with the alginate matrix and provide the alginate microcapsules with the same advantages as chitosan (SMIDSROD and SKJAK-BRAEK, 1990; CHAMPAGNE ET AL., 1992; LARISCH ET AL., 1994). PLL multilayer generation on the alginate microcapsules has been investigated: the first layer of PLL on the microcapsule surface produces a positive charge, so a second alginate layer promotes the formation of a negative layer. This process can be repeated several times. As a result, alginate and PLL layers would be alternately formed (CHAMPAGNE ET AL., 1992; LARISCH ET AL., 1994; MARX, 1989).

Applications and advantages of microencapsulation probiotic microorganisms can be discussed from different angles. Microencapsulations can be used efficiently for the preparation of high viability bacterial starter cultures. The shelf life of Lactobacillus rhamnosus which is kept at room temperature and high humidity is shown to be around six months. This time was significantly increased to 18 months when microorganisms were microencapsulated and maintained at the temperature of liquid nitrogen. Microcapsules containing microorganisms can be added directly to the consumed products. Only 10% deterioration of microcapsules was detected after being subjected to simulated gastrointestinal tract conditions (MATTILA-SANDHOLM ET AL., 2002). Another advantage of microencapsulation of probiotic microorganisms is related to viability in the gastrointestinal tract. Several articles in the literature confirm the efficiency of microencapsulation in increasing the viability of probiotics when they are subjected to the acid - enzymatic conditions of the gastrointestinal tract. As an example, RAO in 1989 demonstrated that microencapsulation of B. pseudolongum with phthalate acetate decellulose increases its viability under conditions simulating gastrointestinal tract (GROBOILLOT ET AL., 1993). Experiments from LEE and HEO (2000) showed that the high survival of calcium alginate microencapsulated B.Iongum under simulated conditions of gastric juice (pH = 1.5) was considerably increased. Experiments have indicated that a calcium chloride coating over the calcium acid alginate microcapsules containing L acidophilus increases microorganism tolerance against extreme acid values (pH = 2) (CHANDRAMOULI ET AL., 2004). Simulation of stomach conditions (pH = 1.5) led to loss of viable cell counts of B. infantice, however, the viability does not exceed 0.67% of the initial viable cell count (SUN and GRIFFITHS, 2000). Research has shown that resistant starch is an efficient component for probiotic micro-encapsulation, as this component is not dissolved or decomposed in gastric juice, at neutral pH and by the enzymatic activity of the pancreas, but releases microorganisms when the microcapsules come into contact with intestinal conditions (ENGLYST ETAL). , 1992; SUN and GRIFFITHS, 2002). An important point to note is that in addition to the type of microcapsule material, the diameter of the microcapsules or the coating of the capsules is a determining factor in maintaining the viability of probiotic microorganisms. Excessive reduction in diameter may weaken or even remove the protection function of the microcapsules (SULTANA ET AL., 2000). The ideal diameter range depends on the microorganism to be immobilized.

Another application for microcapsules containing probiotic microorganisms is the production of biomass and industrial fermentations. Thus, microencapsulation of microorganisms may include the following applications: increased tolerance of microorganisms to factors such as bacterial infection (STEESON ETAL., 1987), poisonous chemical agents, protection of microorganisms against mutagenic agents, thus achieving good productivity in the production of metabolites especially. at high agitation rates (ARNAULDET AL., 1992) and a denser biomass production (CHAMPAGNE ETAL., 1992).

The production of probiotic products is another field of application of microcapsules. The advantages of probiotic microencapsulation in probiotic food products can be discussed from four factors: 1) increased viability of probiotics in products up to the time of consumption; 2) realization of a new method in food production; 3) fixation and increase of sensory properties of products.

1) Microencapsulation may notably improve the viability of probiotic microorganisms due to its protective effects against high acidity environment factors, low pH, presence of molecular oxygen (in the case of anaerobic microorganisms), harmful agents generated during the process (especially heat treatments). , digestive enzymes, bacteriophages, hydrogen peroxide, short chain fatty acids, carbonyl aromatic compounds (MORTAZAVIAN ET AL., 2006). Increasing the viability of probiotic microorganisms will consequently result in increased shelf life of the product. Undoubtedly, the high acidity and low pH of fermented products are the main factors that cause the lost viability of probiotic microorganisms, especially during refrigerated packaging (SHAH ET AL., 1995; DAVE and SHAH and LANKAPUTHRA, 1997; MORTAZAVIAN ET AL., 2006 ). The microencapsulation of L. acidophilus and bifidobacteria in a calcium alginate matrix does not significantly increase the viability of the microorganisms following their extreme acidity conditions (pH = 2), but under milder acidic conditions (natural yoghurt acidity) observed a microorganism survival. probiotics for more than eight weeks in refrigerated stock. HACS (high ammonia cornstarch) or alginate-RS (resistant starch) mixtures compared with calcium alginate alone improves the morphology of the formed microcapsules with regard to consistency and structural continuity and consequently the viability of microorganisms increase (SULTANA ET AL., 2000). Experiments performed by KEBARY ET AL. (1998) showed that microencapsulation of bifidobacteria with alginate could significantly increase its viability in frozen milk, while using k-carrageenan showed no change. B. Iongum microencapsulated in meiolaceous showed higher viability compared to free microorganisms during the same storage time (TRUELSTRUP-HANSEN ET AL., 2002). According to the investigations carried out by KALIL and MANSUR (1998), the microencapsulation of Bifidobacterium spp. with calcium alginate significantly the viability of microorganisms in mayonnaise with pH around 4.4. The average size of the microcapsules was 3mm after the microencapsulation process (SUN eGRIFFITHS, 2000). Microencapsulated probiotics with the alginate-starch mixture and microcapsule size between 0.5 to 1.0 mm were considerably more viable in yogurt during the storage period (SULTANA ET AL. 2000). Lactobacilli viability was increased in dairy ice cream after alginate microencapsulation (size 25 to 62μιτι) was presented (SHEU and MARSHALL, 1993). The same results were obtained for frozen and fermented desserts. Recovering alginate microcapsules with PLL (PoIi-L-Lysine) has considerably increased viability against severe process conditions (SHAH and RARU LA, 2000). Other researchers indicated that survival of Sifidobacteriumspp. and L. acidophilus significantly increase in frozen fermented desserts when alginate with SPS eTween 80 additives were used for microencapsulation (SULTANAET AL., 2000). Improvement of viability of B. bifidum in yogurt following calcium alginate microencapsulation after three weeks stored under refrigeration at 4 ° C is supported by the counting of viable microorganisms not below 107UFC / mL. In addition, no undesirable sensory properties were detected in the final product. The above mentioned results were also obtained after stocking of frozen product (SULTANA ET AL., 2000). It has been observed that the viability of lactobacillus microencapsulated with calcium alginate could be increased by over 40% in frozen products such as ice cream and frozen milk (SHEU and MARSHALL, 1993). Because the microencapsulation of probiotic microorganisms considerably decreased their metabolic activity, the viability of the microorganisms should increase due to the acid production rate. Therefore, it was shown that the incubation time for yoghurt made with L. casei and L. acidophilus above the pH 5 endpoint increased from 6 hours for free microorganisms to 30 hours for microencapsulated microorganisms (SULTANA ETAL., 2000). The decrease in acidification rate of bacteria and as a result of the drop in pH decrease in this period leads to a considerable increase in shelf life of the product due to the increase in the viability of microorganisms within the storage time (MORTAZAVIANET AL., 2006).

2) Using the microencapsulation process early cultures of probiotic microorganisms innovations have been developed in the production of dairy probiotic products as well as yogurt. Microencapsulation of probiotic microorganisms may cause a desirable rate of metabolic cellular activity. For example, new continuous methods of producing yoghurt with traditional microencapsulated probiotic bacteria (Streptococcus salivarius ssp. Thermophilus and Lactobacillusdelbrueckii ssp. Bulgaricus) have been proposed and have the following advantages compared to traditional methods: products with relatively constant sensory properties can be obtained. The viability of the remaining bacteria is much higher and the proportion of Lactobacillus delbrueckii ssp.bulgaricus / Streptococcus thermophilus from the early stage to the end of the fermentation process can be controlled as well as the organoleptic properties of the product (KRASAEKOOPT ET AL., 2003).

As disadvantages of producing fermented products using microencapsulated starter cultures we can mention the long incubation period and high prices due to the need to use high amounts of inoculum (due to the lack of cell multiplication during the fermentation process) can be mentioned. However, according to Larisch et al (1994) the incubation time of Lactococci, microencapsulated with PLL1-coated alginate decreases by a factor of 17% compared to the conditions of yogurt fermentation with free microorganisms. However, there is no further record to confirm such observation.

The microencapsulation of probiotic microorganisms has been used to produce a homogeneous dispersion of microorganisms within the product. This is very important mainly in viscous products having more than one phase (polyphase) as well as mayonnaise (KHALIL and MANSOUR, 1998).

Microencapsulation of probiotic microorganisms helps to maintain or improve the sensory properties of the final product. In general, fermented products (as well as yogurt) produced by encapsulated microorganisms are milder compared to those produced by unencapsulated microorganisms due to the lower amount of acids produced (ADHIKARI ET AL., 2000). Consequently, microencapsulation of early cultures promotes an aromatic fixation of fermented products as encapsulated cells are relatively or totally inactive of metabolism and do not influence the aroma of the products, especially during storage time. As an example, no significant variation in sensory properties of B. bifidumencapsulated yoghurt containing yoghurt was observed after three weeks of refrigerated storage at 4 ° C (KRASAEKOOPT ET AL., 2003).

Although microencapsulation of probiotic microorganisms can be applied as an efficient method to improve the sensory properties of probiotic products, their improper use can lead to a drop in such properties and texture of the final product, especially taste defects. This sensation is related to microcapsule size (TRUESTRUP). (Hansen et al., 2002; Chandramuli et al., 2004).

Probiotic drinks are functional foods, that is, besides providing basic nutrition, they promote health. These foods have the potential to promote health through unpredictable mechanisms through conventional nutrition, and it should be noted that this effect is restricted to health promotion and not cure of disease (Sanders, 1998). The human gastrointestinal tract is a kinetic microecosystem that enables the normal performance of host physiological functions unless epotentially pathogenic harmful microorganisms dominate. Maintaining an appropriate balance of the microbiota can be ensured by systematic supplementation of the diet with probiotics, prebiotics and symbiotics (BIELECKA, BIEDRZYCKA, MAJKOWSKA, 2002). In light of this fact, in recent years, the concept of functional foods has come to focus intensively on food additives that may have a beneficial effect on the composition of the intestinal microbiota (ZIEMER, GIBSON, 1998).

Probiotics were classically defined as food supplements based on living microorganisms, which beneficially affect the host animal, promoting its intestinal microbiota balance (FULLER, 1989). Several other definitions of probiotics have been published in recent years (Sanders, 2003). However, the definition currently accepted internationally is that they are living microorganisms, administered in adequate amounts, that confer host health benefits (FOOD AND AGRICULTURE ORGANIZATION OF UNITED NATIONS; WORLD HEALTHORGANIZATION, 2001; SANDERS, 2003).

The beneficial influence of probiotics on the human microbiotestinal includes factors such as antagonistic effects, competition and immunological effects, resulting in increased resistance to pathogens. Thus, the use of bacterial probiotic cultures stimulates the multiplication of beneficial bacteria, in detriment to the proliferation of potentially harmful bacteria, reinforcing the natural mechanisms of host defense (PUUPPONEN-PIMIA ET AL., 2002).

Under normal conditions, numerous species of bacteria are present in the gut, most of them strict anaerobic. This composition makes the gut capable of responding to possible anatomical and physicochemical variations (LEE ET AL., 1999). Intestinal microbiota exerts considerable influence on a series of biochemical reactions of the host. At the same time, when in balance, it prevents potentially pathogenic microorganisms present in it from exerting their pathogenic effects. On the other hand, the unbalance of this microbiota may result in the proliferation of pathogens, with consequent bacterial infection (ZIEMER, GIBSON, 1998).

Healthy microbiota is defined as the microbiotanormal that conserves and promotes well-being and absence of disabilities, especially of the gastrointestinal tract. The correction of the properties of the unbalanced autochthonous microbiota constitutes the rationality of probiotic therapy (ISOLAURI, SALMINEN, OUWEHAND, 2004). The beneficial influence of probiotics on human intestinal amicrobiotic includes factors such as antagonistic effects and competition against undesirable microorganisms and immunological effects (PUUPPONEN-PIMIÃ ET AL., 2002). Experimental data indicate that several probiotics are able to modulate some characteristics of digestive physiology, such as mucosal immunity and intestinal permeability (FIORAMONTI, THEODOROU, BUENO, 2003). The binding of probiotic bacteria to enterocyte cell surface receptors also initiates the cascade reactions that result in cytokine synthesis (KAUR, CHOPRA, SAINI, 2002).

The knowledge of the intestinal microbiota and its interactions led to the development of eating strategies, aiming at maintaining and stimulating the normal bacteria present (GIBSON, FULLER, 2000). It is possible to increase the number of health-promoting microorganisms in the gastrointestinal tract (GIT) by introducing probiotics by feeding or by consuming prebiotic food supplements, which will selectively modify the composition of the microbiota, giving the probiotic a competitive advantage over other ecosystem bacteria ( CRITTENDEN, 1999).

Three possible mechanisms of action are attributed to probiotics, the first of which is the suppression of the number of viable cells through the production of compounds with antimicrobial activity, competition for nutrients and competition for adhesion sites. The second of these mechanisms would be the alteration of microbial metabolism by increasing or decreasing enzymatic activity. The third would be the stimulation of host immunity through increased antibody levels and increased macrophage activity. The spectrum of activity of probiotics can be divided into nutritional, physiological and antimicrobial effects (FULLER, 1989).

The health benefits of the host attributed to the ingestion of the most prominent probiotic cultures are: control of the intestinal microbiota; stabilization of the intestinal microbiota after antibiotic use; promotion of gastrointestinal resistance to pathogen colonization; decrease of the pathogen population through the production of acetic and lactic acids, bacteriocins and other antimicrobial compounds; promotion of dalactose digestion in lactose intolerant individuals; immune system stimulation; constipation relief; increased absorption of minerals and vitamin production. Although not yet proven, other effects attributed to these cultures are decreased risk of colon cancer and cardiovascular disease. Decreased plasma cholesterol concentrations, antihypertensive effects, reduced ulcerative activity of Helicobacter pylori, control of rotavirus and Clostridium difficile-induced colitis, prevention of urogenital infections, and inhibitory effects on mutagenicity (SHAH, LANKAPUTHRA) are also suggested. , 1997; CHARTERIS ET AL., 1998; JELEN, LUTZ, 1998; KLAENHAMMER, 2001; KAUR, CHOPRA, SAINI, 2002; TUYY ETAL., 2003).

Among the existing types of probiotics, we highlight Kefir, a name given to both grains and beverages resulting from fermentation. Kefir originates from the Caucasus, consisting of a colony containing bacteria and yeast, bound together by a complex structure of proteins, lipids and carbohydrates. There are two types of Kefir grains: 1) Milk - because it fermentes various types, producing a drink similar to eco-yogurt; 2) Water - Fermentes a solution with water, sugar elimão, which results in a refreshing carbonated drink. The exact combination of bacteria and yeast varies between these two types of grain, but they have more than 50 types of microorganisms. Being a colony with several types of probiotic microorganisms, the drinks obtained from these grains are also probiotic drinks.

From the study of the fermentative kinetics of these grains were produced other fermented beverages with various substrates, such as fruit juices, coconut water, soy extract, which we call Kefir BioLogicus.

Probiotics, as well as Kefir BioLogicus have a very laborious handling, since care is needed for the preservation of microorganisms (consortium of microorganisms), such as the regular exchange of culture medium, the (also regular) reception of kefir grains formed in the culture. and conservation in a certain temperature range. In addition, both kefir and probiotic microorganisms are generally not intended to enrich non-probiotic foods and cosmetics, as their strong taste and core odor characteristics endanger the "enriched" food, making it undesirable. One way to avoid such a problem is the microencapsulation process.

There are several techniques that can be used for microencapsulation, and the method selection is dependent on the application that will be given to the microcapsule, the desired size, the release mechanism and the physicochemical properties of both the active material and the encapsulating agent ( JACKSONe LEE, 1991). Microcapsule sizes may range from a few nanometers to several micrometers; The form is also quite variable depending on the method and encapsulating agent used to prepare them.

The choice of encapsulating agent depends on a number of factors, including non-reactivity with the material to be encapsulated, the process used for microcapsule formation and the optimal release mechanism. Many materials can be used as a cover for microcapsules, such as gum arabic, alginate Agar1 and carrageenan; carbohydrate starch, modified starches, dextrins and sucrose; cellulosescarboxymethylcelluloses, acetylcellulose, nitrocellulose; lipids, paraffin, mono- and diaglycerols, oils and fats; inorganic materials calcium sulfate and silicates; gluten, casein, gelatin and albumin proteins (JACKSON and LEE, 1991).

The release mechanisms of encapsulated active materials vary according to the nature of the encapsulating agentemic, and they usually occur due to mechanisms such as temperature and pH variation, media solubility, biodegradation, diffusion, mechanical rupture, selective permeability and concentration gradient in relation to release (BAKAN, 1973; BRANNON-PEPPAS, 1993). It should be noted that the thickness of the microcapsule cover can be modified so that stability and permeability are changed (BAKAN, 1973).

The general purposes of microencapsulation may be some of the following: transforming a liquid into solid to facilitate its handling, transport and addition into formulations, separating reactive materials; reduce toxicity of active material, promote controlled release of encapsulated active; reduce volatility or flammability of liquids; mask certain flavor and odor components; increase shelf life; and protect against light, moisture and heat (JACKSON and LEE, 1991).

Materials that may be encapsulated for application in the food industry include acids, bases, oils, vitamins, salts, gases, amino acids, essential oils, dyes, enzymes and microorganisms (DESAI and PARK, 2005).

Microorganisms have been microencapsulated or immobilized to enable their reuse in the production of lactic acid and fermented dairy products (T1 PAYANG and KOZAKI1 1982; HYNDMAN ET AL., 1993; GROBOILLOT ET AL., 1993), to increase cell concentration in reactors, with consequent increase in productivity (YOO ET AL., 1996), to protect them against the presence of oxygen (KIM and OLSON, 1989), against low freezing temperatures (SHEU and MARSHALL, 1993; SHEU ET AL., 1993), against the bactericidal effect of gastric juice and other acid media (RAO ET AL., 1989; MOLDER and VILLA-GARCIA, 1993; DINAKAR eMISTRY, 1994; KHALIL and MANSOUR, 1998; CUI ET AL., 2000; FAVARO-TRINDADE and THICK 2000; SULTANA ET AL., 2000; FAVATO-TRINDADE AND GROSSO, 2002; HANSEN ET AL., 2002; DESMOND ET AL., 2002; PICOT and LACROIX, 2004; IYER eKAILASAPATHY, 2005; MUTHUKUMARASAMY ET AL., 2006; CHEN ET AL., 2006; OLIVEIRA ET AL., 2007; OLIVEIRA ET AL., 2007), to remove them from the product, inte disrupting acidification (CHAMPAGNE and CÔTE, 1987) to increase stability and maintain viability of culture during product storage (KIM ET AL., 1988; CHAMPAGNE ET AL., 1994; Hansen et al., 2002; KAILASAPATHY, 2006; CAPELA ET AL., 2006; MUTUKUMARASAMY and HOLLEY, 2006) and to increase the shelf life of Pseudomonas fluorescens-putida (AMIET-CHARPENTIER ETAL., 1998).

Although microencapsulation is a very innovative technology limited only by the imagination, it is still very little commercially exploited in the area of food.

A variety of methods of immobilizing a highly concentrated culture of microorganisms has been developed to increase productivity in the fermentation process.

One of the most widely used methods of immobilization is the cell entrapment method, where microorganisms are trapped in microcapsule-shaped gels formed of calcium carcinate, polyacrylamide, carrageenan, agarose, gelane, or polytetrafluoroethylene (PTFE) (according to US Pat. No. 5, 175.093; 5 288.632; 5.093,253; 4,722,898; 4,828,997; 5070,019; GHOSH, S., 1998).

However, the cell trapping method, although simple, has shown limitations in increasing the concentration of microorganisms per unit volume of the matrix, because microorganisms can only grow on the surface or interstitial spaces of the matrix.

On the other hand, the microencapsulation method has been applied in different ways for immobilization of animal cells, plants, bacteria, algae or fungi (according to: USPat. No. 5, 286,495; four 806,355; 4,689,293; LIM, F. ET As described above, the development of a method of immobilizing simpler and more effective microorganisms is required.

The fermentation of Kefir BioLogicus is of the lacto-alcoholic type, with the production of several types of acids (carbonic, butyric and acetic), predominantly lactic acid in lactose, derived from milk lactose. Amino acids such as valine, leucine, lysine and serine form during fermentation, plus appreciable amounts of pyridoxine, vitamin B12, folic acid and biotin. Several bioactives are responsible for the functional and probiotic activity of Kefir BioLogicus: the microorganisms themselves (dead or alive), the metabolites of the microorganisms formed during fermentation (antibiotics, including bactericides), food matrix breakdown products such as peptides (OUWEHAND and SALMINEN1998; FARNWORTH 2002).

Kefir's secondary metabolism gives rise to differentiated structures with polysaccharides produced by a variety of lactic acid bacteria including lactobacilli and streptococci - Lactococcus and Leuconostoc (DE VUYST and DEGEEST 1999; RUAS-MADIEDO ET AL. 2002.). These cell surface carbohydrates impart protective and adaptive characteristics to their bacterial producers; and since they are freely bound to the cell membrane, they are easily lost in the environment. In food products, exopolysaccharides generally contribute to the ethanol-optical and stability characteristics. Among these polysaccharides, KEFIRANO stands out, which contains D-glucose and D-galactose in a 1: 1 ratio. Hydrolysis reactions followed by NMR analysis have been used to determine the chemical structure of kephyran. The proposed structure is a hexa or a hepta saccharide which repeats the unit which is itself composed of a regular pentasaccharide unit to which one of the sugar residues bind randomly (KOOIMAN, 1968; MICHELI ET AL., 1999).

Kefirano has been the subject of several studies conducted by BioLogicus in recent years. Scientific papers have been published internationally, which report the great importance of kefiran for the industry in general and, more particularly, for the pharmaceutical industry, as it has proven antibacterial, antimycotic and antitumor activity (JA PIERMARIA ET AL., 2009; RODRIGUES ET AL., 2005; MAEDA ET AL., 2004). There is also research that highlights the possibility of anti-inflammatory and antiallergic effects of lower airways (KWON ET AL., 2009).

Studies with kefirane have shown several beneficial health effects, including suppression of increased blood pressure (MAEDA ET AL., 2004); stress reduction, since they present activities on the production of beta-interferon, cortisol and noradrenaline (S. KBayama et al., 1997), increased phagocytic activity of epulmonary peritoneal macrophages (CG VIDEROLA ET AL., 2005; CG VIDEROLA ET .AL. 2006) and increase of IGA cells at these sites (CG VIDEROLAET AL., 2006); anti-tumor activity (J.R. LIU ET AL., 2002; M.MUROFUSHI ET AL., 1983; M. SHIOMI ET AL. 1982); antimicrobial activity (K.L. RODRIGUES ET AL., 2005); preventive effect on antibiotic-associated diarrhea, by favoring normal intestinal bloom, protecting it against exogenous pathogens and maintaining balance (M. ZUBILLAGA ET AL. 2001); increased activity of intestinal dipeptidase (E. URDANETA ET AL., 2007), reduction of blood lipids, blood pressure, blood glucose and constipation (H. MAEDA ET AL., 2004).

Kefiran is an extracellular polysaccharide produced during the fermentation of Kefir BioLogicus colony, constituting 24 - 25% (w / w) of the dry weight of Kefir BioLogicus grains and is a matrix of fibrillar amorphous material. This matrix surrounds the bacteria and yeast in the kefir BioLogicus grains and holds them together providing protection against microorganisms outside the colony. Structurally kefirane is formed by a branched glucogalactan, consisting approximately of equal amounts of D-glucose and D-galactose residues.

In a recent study by RODRIGUES ET AL., (2005), kefiran inhibited the growth of seven bacterial strains and one pathogenic yeast strain. It has demonstrated tumor activity by inhibiting the growth of solid tumors, carcinoma and sarcoma, an aspect first reported by Shiomu (Shiomuet Al., 1982). This biopharmaceutical also stands out for its anti-metastasis activity against lung carcinoma and melanoma, which has already been described in rats (FURUKAVA, 2001).

Due to its antitumor, antibacterial and antifungal properties, Kefiran can be used as a good healing, anti-inflammatory and antimicrobial agent for use in a wide range of infections (SALOFF-COSTE, 1996).

One of the agents responsible for the properties of dokefirane are bacteriocins, toxins produced by kefiran-beneficial bacteria to inhibit the growth of other similar bacteria as a means of protection.

In addition to the above properties related to the kefiran polysaccharide, which is a byproduct of the secondary metabolism of Kefir BioLogicus, a very important property for the food and cosmetic industry is the use of same as a food additive. This feature is closely related to its properties of being used as a stabilizer in food and cosmetics. According to RIMADAE ABRAHAM (2006), kefiran can be used as a food additive for fermented foods increasing the rheological properties of the product. According to PIERMARIA ET AL (2008), Kephyran is considered a functional additive.

The property of kefirane in forming gels was also a scientific work (PIERMARIA ET AL. 2008) indicating the potential of this exopolysaccharide to be used as a microencapsulation matrix component as well as sodium alginate and other polymeric compounds.

Under the circumstances described above, the present invention presents the development of a simple and effective method regarding the immobilization of a consortium of microorganisms present in Kefir BioLogicus and its bioactive products (proteins, vitamins, among others) employing calcium alginate modified capsules with kefirane pectinae. In contrast to existing methodologies, the present invention discloses the formation of pectin and kefirano modified calcium alginate microcapsules which have the quality of trapping a large amount of microorganisms making them easier to handle for use in the most effective fermentation process. Mixing dealginate with Pectin and / or Kefiran results in the formation of denser and more mechanically resistant microcapsules, thereby improving the protective effect of probiotic microorganisms, thereby improving their viability.

In accordance with the present invention, the inventors have prepared microcapsules based on modified calcium alginate with Kefiran and pectin containing within them a consortium of microorganisms present in Kefir BioLogicus as well as its bioactive products. The consortium of microorganisms present in Kefir BioLogicus is composed of four groups, as described below: 1- Lactobacilli: Lactobacillus acidophilusLactobacillus bulgaricusLactobacillus brevisLactobacillus caseiLactobacillus casei ssp. lactosus; Lactobacillus casei ssp. rhamnosusLactobacillus cellobiosisLactobacillus delbrueckiiLactobacillus fermentumLactobacillus fructivoranLactobacillus helveticusLactobacillus hilgardiiLactobacillus johnsoniiLactobacillus kefir, Lactobacillus kefiranofaciensLactobacillus kefirgranumLactobacillus IaetisLactobacillus paraeaseiLactobacillus parakefirLactobacillus plantarumLactobacillus rhamnosusLactobacillus viridescens

2-Lactococcus / Streptococcus: Enteroeoceus duransLactoeoceus Iaetis subsp. eremorisLactoeoceus Iaetis subsp. IaetisLe. Iaetis var. diaeetylaetisLeueonostoe eremorisLeueonostoe kefirLeueonostoe sp.

S. Iaetis

S. sallivarius ssp.thermophilusStreptoeoeeus thermophilus

3-Yeast: Candida pseudotropiealisC. raneens

Candida kefirC. tenuisK bulgarieusK. fragilisKluyveromyces IaetisKluyveromyces marxianus var. MarxianusTorula kefir

Torulaspora delbrueekiiSaccharomyces boulardiiSaccharomyces earlbergensisSaccharomyees eerevisiaeSaccharomyees IaetisSaccharomyees kefírSaccharomyees eerevisiae

Saccharomyees delbrueekii or Saccharomyees unisporus

4- Molds

Geotriehum eandidum

5- Acetobacter: Aeetobaeter aeetiAcetobacter rasensAcetobacter sp.

6- Other BacteriaBacillus sp.Micrococcus sp.Bacillus subtilis

Given the above scenario, the process presented here is a quick and inexpensive solution for the conservation of the consortium of microorganisms and their bioactive as well as for the enrichment of non-probiotic and cosmetic foods.

The immobilization of the consortium of microorganisms as well as their metabolic products, as performed in the invention described herein, occurs as follows: the consortium of microorganisms and their metabolic products are added to a solution of Kefirano1 sodium alginate and pectin, forming a new solution (solution A ). Solution A can be represented as follows: (100-X-Y)% Alginate - X% Kephyrane - Y% Pectin (by volume percentage). Since the concentration of Sodium Alginate solution may vary from 1 to 5% (by mass), that of Kefiran may vary from 1 to 3% (by mass) and Pectin may vary from 1 to 4% (by mass).

This new solution is dripped into a second Calcium Chloride solution (solution B), the latter of which may range from 0.1 to 4.0% (by mass). The dripping of Ana solution B causes the formation of microcapsules containing the consortium of microorganisms, as well as their metabolic products.

Microcapsules are a practical way of storing, and applying the consortium of microorganisms. they do not require constant maintenance of the Kefir BioLogicus microorganism consortium and products may be added to make them probiotic without transforming their taste, aroma or color, thus overcoming the disadvantages of handling the probiotics and their organoleptic characteristics.

In addition to the present description in order to gain a better understanding of the features of the present invention and in accordance with the preferred practical application thereof, the description accompanies a drawing in which, by way of example, but not limitation, represented the following:

Figure 1 shows that the polymeric solution (solution A, formed by the pool of microorganisms plus kefiran, and / or pectin and sodium alginate) is prepared in tank 1 and stirred to feed tank 2.

From tank 2 the solution is aspirated and dosed by the bombaperistaltic, dripping into the microcapsule formation, notably Ill / 1, which contains the calcium chloride solution (solution B). Once the production of a certain amount of microcapsules is completed, conclusion which may be for time or volume, this tank will be replaced by tank 111/3, which will contain only calcium chloride solution.

The microcapsules in the 111/1 tank, which has been replaced, will be allowed to stand for 5 to 20 minutes, depending on the substrate in which the pool is needed to complete the process.

After this time of rest, the microcapsules and the solution will be poured into a sieve, where there will be the separation of the liquid and solid phases, leaving the liquid phase in tank III / 2, which will replace the receiving tank of bombaperistá Itica droplets.

On the other hand, the collected microcapsules will be taken to tank IV, where, without being removed from the sieve, they will be properly washed with the closed circuit mineral water, thus concluding a production cycle.

According to the above illustration, the present invention refers to an "INDUSTRIAL PROCESS OF MOBILIZING A CONSORTIUM OF MICRORGANISMS, ORIGIN OF KEFIR BIOLOGICUS, AS WELL AS ITS SEBIOACTIVES, THROUGH CODE DEFORMATION OF MICROPHALLATION, In other words, an industrial process that allows the manufacture of microcapsules from a consortium of microorganisms that do not suffer from the limitations inherent to other probiotic foods: they do not require laborious maintenance nor do they influence the taste, aroma or color of the products to which they are added, but retain all the inherent benefits of microorganism consortium (probiotic culture with about 50 types of microorganisms).

The immobilization of the consortium of microorganisms as well as their metabolic products, as performed in the invention described herein, occurs as follows: the consortium of microorganisms and their metabolic products are added to a solution of Kefirano1 sodium alginate and pectin, forming a new solution (solution A). , this solution is kept under stirring until the moment of use (1). Solution A can be represented as follows: (100-X-Y)% Alginate - X% Kephyrane - Y% Pectin (by volume percentage). Since the concentration of Sodium Alginate solution can vary from 1 to 5% (by mass), Kefirano can vary from 1 to 3% (by mass) and Pectin may vary from 1 to 4% (by mass).

At the time of use (2), solution A is transferred to tank 2 where it is aspirated and dosed by a peristaltic pump, which drips into a second solution (solution B), the latter of concentration of Calcium chloride which may vary from 0, 1 to 4.0% (by mass).

Once the production of a certain amount of microcapsules (3) has been completed, which may be either time or volume, this tank will be replaced by tank III / 3, which will contain only calcium chloride solution and will receive drip from the pump. peristaltic.

The microcapsules in the 111/1 tank, which has been replaced, will be allowed to stand for 5 to 20 minutes, depending on the substrate in which the pool of microorganisms is contained, necessary to complement the microencapsulation process. After this rest time, the microcapsules and the solution will be poured into a sieve, where the liquid and solid phases will be separated, leaving the liquid phase in the 111/2 tank, which will replace the bombaperistaltic droplet receiving tank (4) and repeat the process for the production of more microcapsules. .

On the other hand, the collected microcapsules will be taken to tank IV (5), where, without being removed from the sieve, they will be properly washed with the distilled water in closed circuit, concluding with this operation a production cycle of the microcapsules containing the consortium of microorganisms.

Microcapsules form a practical way to store, and apply the consortium of microorganisms: they do not require constant maintenance of the microorganisms and can be added to products to make them unprobiotic without transforming their taste, aroma or color, thus overcoming the disadvantages. handling of probiotics and their organoleptic characteristics.BIBLIOGRAPHIC REFERENCES

ADHIKARI, K .; MUSTAPHA, A .; GRUN, I.U; FERNANDO, L.Viability of microencapsulated bifidobacteria in set yogurt during refrigerated storage. J. Dairy I know. 83: 1946-1951, 2000.

AMIET-CHARPENTIER, C .; GADILLE, P .; DIGAT1 B .; BENOIT1J. P. Microencapsulation of rhizobacteria by spray-drying: formulation and survival studies. Journal of Microencapsulation, London, v. 15, no. 5, 639-659, 1998.

ARIMOTO, M .; ICHIKAWA, H .; FUKUMORI, Y.Microencapsulation of water-soluble macromolecules with acrylicterpolymers by the Wurster coating process for colon-specificdrug delivery. Powder Technology, Lausanne, v. 141, no. 3, 177-186, 2004.

ARNAULD, J.P .; LAROIX, C .; CHOPLIN, L. Effect of agitationrate on cell release rate and metabolism during continuousfermentation with entrapped growing Lactobacillus casei subsp.casei. Biotech Teeh., 6: 265, 1992.

ARSHADY, R. Microcapsules for food. Journal of Microencapsulation, London, v. 10, no. 4,413-435, 1993.

AUDET, P .; PAQUIN, C .; LACROIX, C. Immobilized growinglactic acid bacteria with k-carrageenan-locust bean gum gel.Applied Microbiologyand Biotechnology, v. 29, 11-18, 1988.BAKAN1 J. A. Microencapsulation of foods and related products. Food Technology, v. 27, no. 11, 34-44, 1973.

BEAL, C .; SKOKANOVA, J .; LATRILLE, E; MARTINN, N; CORRIEU, G. Combined effects of culture conditions and time on acidification and viscosity of stirred yogurt. JDairy I know. 82: 673-681, 1999.

BIELECKA, M .; BIEDRZYCKA, E .; MAJKOWSKA, A. Selectionof probioties and prebioties for synbiotics and confirmation oftheir in vivo effectiveness. Food Res. Int., V.35, no.2 / 3, 125-131,2002.

BRANNON-PEPPAS, L. Controlled release in the food andeosmeties industries. In: Polymeric delivery systems: properties and applications. Oxford Oxford University Press, 1993. chapter 3, 42-52.

BUCK, L.M .; GILLILAND, S.E. Comparisons of freshly isolated strains of Lactobacillus acidophilus of human intestinal origin for the ability to assimilate cholesterol during growth. J Dairy Sci., V. 77: 2925-2933, 1994.

CAPELA, P .; HAY, Τ. K. C .; SHAH, Ν. P. Effeet of cryoprotectants, prebioties and mieroeneapsulation on survival of probiotic organisms in yoghurt and freeze-yoghurt. FoodResearch International, v. 39, no. 2, 203-211, 2006. CHAMPAGNE, C. P .; CÔTE, C. Β. Cream fermentation byimmobilized Iactic acid bacteria. Biotechnology Letters1 v. 9, no. 329-332, 1987.

CHAMPAGNE, C. P .; GARDNER, N .; DUGAL, F. Increasing thestability of immobilized Lactococcus Iactis cultures stored at 4 ° C.J. of lnd. Microbiology, v. 13, no. 6, 367-371, 1994.

CHAMPAGNE, C.P .; GAUDY, C .; PONCELET, D .; NEUFELD, R.J. Lactococcus Iaetis release from alginate beads. ApplEnviron Microbiol., V. 58: 1429-1434, 1992.

CHAMPAGNE, C.P .; MORIN1 N .; COUTURE, R .; GAGNON, C.; JELEN, P .; LACROIX, C. The potential of immobilized cell technology to produce freeze-dried, phage-protect cultures of Laetococeus latis. Food Res Int., V. 25, 419-427, 1992.

CHANDRAMOULI, V .; KAILASAPATHY, K .; PEIRIS, P .; JONES, M. An improved method of microencapsulation and its evaluationto protect Lactobacillus spp. in simulated gastric conditions. Journal of Microbiological Methods, v. 56, no. 27/35, 2004.

CHARTERIS, W.P .; KELLY, P.M .; MORELLI, L .; COLLINS, J.K.Ingredient selection criteria for probiotic microorganisms infunctional dairy foods. Int. J. Dairy Teehnol., V.51, no. 4, 123-136,1998.

CHEN, K.N .; CHEN, M. J .; LIN, C. W. Optimal combination ofthe encapsulating materials for probiotic microcapsules and itsexperimental verification (R1). Journal of Food Engineering, v.

76, no. 3, 313-320, 2006.

CRITTENDEN, R.G. Prebiotics. In: TANNOCK, G.W., ed.Probiotics: A Critical Review. Norfolk: Horizon Scientific Press, 1999. 141-156.

CUI, J. H .; GOH, J. S .; KIM1 P. H .; CHOI, S. H .; LEE, B. J. Survival and stability of bifidobacteria Ioaded in alginate poly-L-Iysine microparticles. International Journal of Pharmaceuticals, v. 210, no. 1-2, 51-59, 2000.

DALY, C .; DAVIS, R. The biotechnology of lactic acid bacteria with emphasis on application in food safety and human health / agricultural and food science in Finland. Agric Food Sci., V. 7, 251-265, 1998.

DAVE, R. I .; SHAH1 Ν. P. Viability of yoghurt and probiotic bacteria in yoghurts made from commercial starter cultures. International Dairy Journal, v. 7, 31-41, 1997.

DAVE1 R.I .; SHAH, N.P. Evalution of media for selective enumeration of Streptococcus thermophilus, Lactobacillusdelbruekii spp. bulgarieus, Lactobacillus aeidophilus andbifidobaceria. J Dairy Sci., V. 79: 1529-1539, 1996.

DAVE1 R.I .; SHAH1 N.P. Viability of yoghurt and probiotic bacteria in yoghurts made from commercial starter cultures. IntDairyJ., V. 7, 31-41, 1997. DE VUYST, L .; VANDERVEKEN, F .; VEN1 S. VAN; DEGEEST, Β. Production by and isolation of exopolysaccharidesfrom streptoeoeeus thermophilus grown in a milk medium andevidence for their growth-assoiated biosynthesis. Journal of Applied Microbiology, v.84, no. 6, 1059-68, 1998.

DESAI, K. G. H .; PARK, H. J. Recent developments in micronencapsulation of food ingredients. Drying Technology, v.23, no. 7, 1361-1394, 2005.

DESMOND, C .; ROSS, R. P .; OOALLAGHAN, E.; FITZGERALD, G .; STANTON, C. Improved survival of Lactobacillus paracasei NFBC 338 in spray-dried powders containing gum acacia. Journal of Applied Microbiology, v. 93, no. 6, 1003-1011, 2002.

DIMANTOV, A .; GREENBERG, M .; KESSELMAN, E .; SHIMONI.Study of high amylase corn starch as food grade enteric coatingin a mieroeapsule model systems. Innov. Food Sci EngTechnol., V. 5, 93-100, 2003.

DINAKAR, P .; MISTRY, V. V. Growth and viability of Bifidobacterium bifidum in eheddar cheese. Journal of DairyScience, v. 77, no. 10, 2854-64, 1994.

EIKMEIER, H .; REHM, H.J. Stability of calcium-alginate during citric acid production of immobilized Aspergillus niger. ApplMicrobiol Biotechnol., V. 26, 105-111, 1987. Bellenton, J.C. (1998). Encapsulation bifidobacteria. Masterthesis, University of Guelph.

ENGLYST, H.N .; KINGMAN, S.M .; GUMMINGS, J.H.Classification and measurement of nutritionally important starchfractions. Eur J Clin Nutr., V. 2, 46 33-50, 1992.

FAO / WHO Experts' Report. Health and nutritional properties of probiotics in food including powder milk with Iive Iactic acidbacteria, 2001.

FARNWORTH ER. Handbook of fermented functional foods.2002; 113-144.

FAVARO-TRIN DADE, C. S .; GROSSO, C.R. F. Microencapsulation of L. acidophilus (La-05) and B. Iaetis (Bb-12) and evaluation of their survival at pH values of thestomach and in bile. Journal of Microencapsulation, v. 19, no. 485-494, 2002.

FAVARO-TRINDADE, C. S .; GROSSO, C. R. F. The effect of the immobilization of L acidophilus & B. Iactis in alginate on their tolerance to gastrointestinal secretions. Milchwissenschaft, v.55, no. 9, 496-499, 2000.

FIORAMONTI, J .; THEODOROU, V .; BUENO, L. Probiotics: what are they? What are their effects on gut physiology? BestPract Res. Clin. Gastroenterol., V.17, 711-724, 2003.FULLER, R. (1992). Probiotics: the scientific basis. London: Chapman & Hall.

FULLER, R. Probiotics in human medicine. Gut1 v. 32: 439-442,1991.

FULLER, R. Probiotics in man and animals. Journal of Applied Bacteriology, v. 66, 365-378, 1989.

FURUKAWA, N .; MATSUOKA, A .; TAKAHASHI, T.; YAMANAKA, Y. Anti-metastic effect of kefir grain components on Lewis Iung carcinoma and highly metastic B16 melanoma inmice, J Agric Sci, 45, 2000;

GARDINI, F .; LANCIOTTI, R .; GUERZONI, M.E .; TORRIANI, S.Evaluation of aroma production and survival of Streptococcusthermophilus, Lactobacillus delbruekii subsp. Bulgarieus and Lactobacillus aeidophilus in fermented milks. Int Dairy J., v.9,125-134, 1999.

GHOSH, S., Immobilization of Baker Yeast Using Plaster of Paris, Biotechnol. Tech., V.2, no. 3,221-219, 1988.

GIBSON, G.R .; FULLER, R. Aspects of in vitro and in vivoresearch approaches directed toward identifying probiotics andprebioties for human use. J. Nutr., V. 130, 391S-394S, 2000. GILLILAND, S. Ε. Acidophilus milk-products from review of potential benefits to consumers. Journal of Dairy Science, v.72, 2483-2495, 1989.

GILLILAND, S.E .; SPECK1 M.L. Instability of Lactobacillusacidophilus in yogurt. J Dairy Sci., V. 60: 1394-1398, 1997.

GISMONDO, M. R .; DRAGO, L .; LOMBARDI, A. Review of probiotics available to modify gastrointestinal flora. International Journal of Antimicrobial Agents, v. 12, 287-292, 1999.

GOLDIN, B.R. Health benefits of probiotics. British J Nutr., V.80, 203-207, 1998.

GOMES, A.M.P; MALCATA, F.X. Bifidobacterium spp and Lactobacillus acidophilus: biological and therapeutic revalant foruse a probiotics. Trends Food Sci Technol., V.10, 139-157,1999.

GOUIN1 S. Microencapsulation: industrial appraisal of existingtechnologies and trends. Trends in Food Science and Technology, v. 15, no. 7-8, 330-347, 2004.

GROBOILLOT, A. F .; CHAMPAGNE, C. P .; DARLING, G. D. PONCELET, D. Membrane formation by interfacial croos-linkingof chitosan for microencapsulation of Laetococeus laetis.Biotechnology and Bioengineering, v. 42, no. 10, 1157-1163,1993.GURR1 M.I. (1987). Nutritional aspects of fermented milkproducts. Milk: The Vital Force Proceeding from the XX11 International Dairy Congress, Dordrecht1 Netherlands1 September29-October 3: 1986.

HAMILTON-MILLER, J.M.T .; SHAH1 S .; WINKLER, J.T. Publichealth issues increased from microbiological and survival in yogurt.Public Health Nutr., V. 2: 223-229, 1999.

HANSEN, L. T .; WONJTAS, A. P. M .; JIW1 Y. L .; PAULSON, A.T. Survival of Ca-alginate microencapsulated Bifidobacteriumspp. In: milk and simulated gastrointestinal conditions. Food Microbiology, v. 19, no. 1, 35-45, 2002.

HOLCOMB, J.E .; FRANK, J.F .; MCGREGOR, J.U. Viability of Lactobacillus acidophilus and Bifidobacterium bifidum insoftserve frozen yogurt. Cult Dairy Product J., v. 26, 4-5, 1991.

HYNDMAN, C.L .; GROBOILLOT, A. F .; PONCELET, D.; CHAMPAGNE, C. P .; NEUFELD, R. J. Microencapsulation of Lactococcus Iaetis within cross-liked gelatin membranes. Journal of Chemistry Technology Biotechnology, v. 56, no. 3,259-63, 1993.

ISOLAURI, E .; SALMINEN, S .; OUWEHAND, A.C. Probiotics.Best Pract. Res. Clin. Gastroenterol., V.18, n.2, 299-313,2004.IWANA1 H .; MASUDA, H .; FUJISAWA, T .; SUZUKI, H.; MITSUOKA, T. Isolation of Bifidobacterium spp. in commercialyoghurts sold in Europe. Bifidobacteria Microflora, v. 12, 39-45, 1993.

IYER, C .; KAILASAPATHY, K. Effect of co-encapsulation of probiotics with prebiotics on increasing the viability ofencapsulated bacteria under in vitro acidic and bile saltconditions and in yogurt. Journal of Food Science, v. 70, no. 1.18-23, 2005.

JACKSON, L. S .; LEE, K. Microencapsulation and Food Industry.LWT - Food Science and Technology, v. 24, no. 4,289-297,1991.

JAN KOWSKI, T .; ZIELINSKA, M .; WYSAKOWSKA.Encapsulation of Iaetie and bacteria with alginate / starchcapsules. Biotechnol Technol., V. 11, 31-34, 1997.

JELEN, P .; LUTZ, S. Funetional milk and dairy products. In: MAZZA1 G., ed. Functional foods: biochemical and processing aspects. Lancaster: Teehnomic Publishing, 1998.357-381 .;

KABAYAMA, S .; OSADA, K .; TACHI, H .; KATAKURA, Y.; SHIRAHATA, S. Enhaneing effects of food components on the production of interferon β from animal cells suppressed by stresshormones, Cytotechnology, v. 23, no. 1-3, 1997.KAILASAPATHY, Κ. Survival of free and encapsulated probiotic bacteria and their effect on the sensory properties of yoghurt.LWT-Food Science and Technology, v. 39, no. 10, 1221-1227,2006.

KAILASAPATHY, K .; RYBKA1 S. L acidophilus and Bifidobacterium spp .: their therapeutic potential and survival inyogurt. Aust J Dairy Technol., V. 52, 28-35, 1997.

KAUR, I.P .; CHOPRA, K .; SAINI, A. Probiotics: potential pharmaceutical applications. Eur. J. Pharm. Sci., V.15, 1-9,2002.

KEBARY, K.M.K .; HUSSEIN, S.A .; BADAWI R.M. Improvingviability of Bifidobacterium and their effect on frozen ice milk. JDairy Sci., V. 26, 319-337, 1998.

KHALIL, A. H .; MANSOUR, Ε. H. Alginate encapsulationbifidobacteria survival in mayonnaise. Journal of Food Science, v. 63, no. 4, 702-705, 1998.

KIM, H. S .; KAMARA, B. J .; GOOD, I. C .; ENDERS Jr., G. L. Method for the preparation of stable microencapsulated Iacticacid bacteria. Journal of Industrial Microbiology, v. 3, no. 4,253-57, 1988.

KIM, H.S .; GILLILAND, S. Lactobacillus acidophilus as a dietary adjunct for milk to aid Iaetosis digestion in human. J Dairy Sci., V.66, 959-966, 1983.KIM1 S. C .; OLSON1 Ν. F. Production of methanethiol in milk fat-coated microcapsules containing Brevibacterium Iinens and Methionine. Journal of Dairy Research, v. 56, no. 5, 799-811.1989.

KIM1 Y. D .; MORR, C. V .; SCHENZ, T. W. Microencapsulationproperties of gum Arabic and several food proteins: Liquidorange oil emulsion particles. Journal of Agricultural Food Chemistry, v. 44, 1308-1313, 1996.

KLAENHAMMER, T.R. Probioties and prebiotics. In: Foodmicrobiology: Fundamentals and Frontiers, DOYLE, M.P.; BEUCHAT, L.R .; MONTVILLE, T.J., 2.ed. Washington: ASM, 2001, 797-811.

KLEIN, G .; PACK, A .; BONAPARTE, C .; REUTER, G. Taxonomy and physiology of probiotic acid bacteria. Int J FoodMicrobiol., V. 41, 103-125, 1998.

Klien, J .; STOCK, J .; VORLOP, K.D. Pore size and propertiesof spherical calcium alginate biocatalysts. Eur J Appl Microbiol Biotechnol., V. 18, 86-91, 1983.

KOOIMAN, P. The chemical structure of kefiran, the water-soluble polysaccharide of kefir grain, Carbohydrate Research, v. 7, 200-211, 1968.

KRASAEKOOPT, W .; BHANDARI, B .; DEETH, H. The influenceof coating materials on some properties of alginate beads and survival of microencapsulated probiotic bacteria.International Dairy Journal, v. 14, 737-743, 2004.

KRASAEKOOPT, W .; BHANDARI, B .; EETH1 H. Evaluation of encapsulation techniques of probiotics for yoghurt. InternationalDairy Journal, v. 13, 3-13, 2003.

KURMAN, J.A .; RASIC, J.L. The Health Potential of Products Containing Bifidobacteria. In: Therapeutic Properties of Fermented Milks, Robinson, RK (Ed.). Elsevier Applied Science, London, UK, pp: 117-157, 1991

KURMANN, J.E. (1983). Bifidobacteria and their role. Basel: Birkhauser. Robinson (Ed). London: Elsevier Application FoodScience series.

LANCAPUTHRA, W.E.V .; SHAH N.P. Bifidobacterium spp in the presence of acid and bile salts. Cult Dairy Prod J., v. 30, 2-7.1995.

LARISCH, B.C .; PONCELET, D .; CHAMPAGNE, C.P.; NEUFELD, R.J. Microencapsulation of Lactococcus Iaetis subsp.Creamoris. J Microencap., V. 11, 189-195, 1994.

LEE, K.I .; HEO, T.R. Survival of Bifudobacterium Iongumimmobilized in calcium alginate beads in simulated gastric juicesand bile salt solution. Appl Environ Microbiol., V. 66, 869-973,2000.LEE, Y.K .; NOMOTO, K .; SALMINEN, S .; GORBACH, S.L. Handbook of probiotics. New York: Wiley, 1999. 211.

LIM1 F .; SUN, A.M. Microencapsulated Islets as BioartifiableEndoerine Pancreas, Science, v. 210, 908-910, 1980.

LIU, J. R .; WANG, S. Y .; LIN, Y. Y .; LIN, C. W .; NUTR ANDCANER, v.44, 183, 2002.

MACFARLANE, G.T .; CUMMINGS, J.H. Probiotics andprebiotics: can regulating the activities of intestinal bacteriabenefit health. BMJ., V. 318, 999-1003, 1999.

MAEDA, H .; ZHU, X .; OMURA, K .; SUZUKI, S .; KITAMURA, S., Effects of an exopolysaccharide (kefiran) on lipids, bloodpressure, blood glucose, and constipation, Biofactors, v. 22,197-200, 2004.

MAEDA1 X .; ZHU, S .; SUZUKI, K .; KITAMURA, S. Structuralcharacterization and biological activities of an exopolysaccharidekefiran produced by Lactobacillus kefiranofaciens WT-2B (T), JAgric Food Chem, v. 52, 5533-5538, 2004.

MARTINSEN, A .; SKJAK-BRAEK, C .; SMIDSROD, O. Alginateas immobilization material: 1. correlation between chemical andphysical properties of alginate gel beads. Biotechnol Bioeng., V. 33, 79-89, 1989. MARX, J.L. (1989). A revolution in biotechnology. Cambridge: Cambridge University.

MATTILA-SANDHOLM1 T .; MYLLARINEN, P .; CRITTENDEN, R .; MOGENDEN, G .; FONDEN1 R .; SAARELA, M.Technological challenges for future probiotic foods. Internationa! DairyJournaI1 v. 12, 173-182, 2002.

MICHELI, L .; UCELLETTI, D .; PALLESCHI, C .; RESCENZI, V. Isolation and characterization of a ropy Lactobacillus strainproducing the exopolysaccharide kefiran. Applied Microbiologyand Biotechnology, v. 53, 69-74, 1999.

MODLER, H. W .; VILLA-GARCIA, L. The growth of Bifidobacterium Iongum in a whey-based medium and viability ofthis organism in frozen yogurt with Iow and high levels ofdeveloped acidity. Cultured Dairy Products Journal, v. 28, no. 4-8, 1993.

MOMBELLI, B .; GISMONDO, M.R. The use of probiotics in medical practice. Int J Ant Agents., V. 16, 531-536, 2000.

MORT AZAVI AN, A.M .; EHSANI, M.R .; MOUSAVI, S.M.; REINHEIMER, J.A .; EMAMDJOMEH, Z .; SOHRABVANDI, S.; REZAEI, K. Preliminary investigation of the combined effect of heat treatment incubation temperature on the viability if the probiotic microorganisms in freshly made yoghurt. Int J DairyTechnol., V. 59, 8-11, 2006. MORTAZAVIAN, Α.Μ .; SOHRABVANDI, S. (2006). Probioticsand food Probiotic products., Eta Publication, Iran.

MORTAZAVIAN, A.M .; SOHRABVANDI, S .; MOUSAVI, S.M.; REINHEIMER, J.A. Combined effects of heating variables on the viability of Probiotic mieroorganisms in yogurt. Aust J DairyTeehnol., V. 61, 248-252, 2006.

MORTAZAVIAN, A.M .; SOHRABVANDI, S .; REINHEIMER, J.A.MRSbiIar agar: Its suitability for the enumeration of mixedprobiotic cultures in cultured dairy products.Milchwissenschaft., V. 62, 270-272, 2006.

MUROFUSHI, M .; SHIOMI, M .; AIBARA1 K. Effect of orallyadministered polysaccharide from kefir grain on delayed-type hypersensitivity and tumor growth in mice. Japanese Journal of Medical Science & Biology, v. 36, no. 1, 49-53, 1983.

MUTHUKUMARASAMY, P .; HOLLEY, R. A. Microbiological and sensory quality of dry fermented sausages containingalginatemicroencapsulated Lactobacillus reuteri. International Journal of Food Microbiology, v. 111, no. 2, 164-169, 2006.

MUTHUKUMARASAMY, P .; WOJTAS, Ρ. THE.; HOLLEY1 R. A. Stability of Lactobacillus reuteri in different types ofmicrocapsules. Journal of Food Science, v. 71, no. 1, 20-24,2006.OLIVEIRA, A. C .; MORETTI, Τ. S.; BOSCHINI, C .; BALIERO, J.C. C .; FREITAS, O .; FAVARO-TRINDADE, C. S. Stability of microencapsulated B. Iactis (BI 01) and L. acidophilus (LAC 4) bycomplex coacervation followed by spray drying. Journal of Microencapsulation, v. 24, no. 7, 685-693, 2007.

OLIVEIRA, A. C .; MORETTI, T. S .; BOSCHINI, C .; BALIERO, J.C. C .; FREITAS, L. A. P .; FREITAS, O .; FAVARITY-TRINITY, C. S. Microencapsulation of B. Iactis (BI 01) and L. acidophilus (LAC 4) by complex coacervation followed by spouted-beddrying. Drying Technology, v. 25, no. 10, 1687-1693, 2007.

OUWEHAND, A.C .; SALMINEN, S.J. Adhesion inhibitory activity of beta-lactoglobulin isolated from infant formulae. Pediatric Act, v. 87, no. 5, 491-493, 2004.

PICOT, A .; LACROIX, C. Effects of micronization on viability and thermotolerance of probiotic freeze-dried cultures. InternationalDairy Journal, v. 13, 455-462, 2003.

PICOT, A .; LACROIX, C. Encapsulation of bifidobacteria in wheyprotein-based microcapsules and survival in simulated gastrointestinal conditions and in yoghurt. International DairyJournal, v. 14, no. 6, 505-515, 2004.

PIERMARIA J. A .; DE LA CANAL, M.L .; ABRAHAM, A. G. Gelling properties of kefiran, the food-grade polysaccharideobtained from kefir grain, Food Hydrocolloids, v.22, n. 8, 1520-1527, 2008. PREVOST, H .; DIVIES, C. Cream fermentation by a mixed culture of Iactococci entrapped in two-layer calcium alginate gelbeads. Biotechnol Let., V. 14, 583-588, 1992.

PUUPPONEN-PIMIÀ, R .; AURA, A.M .; OKSMAN-CALDENTEY, K.M .; MYLLARINEN, P .; SAARELA, M .; MATTILA-SANHOLM, T .; POUTANEN, K. Development of functional ingredients for guthealth. Trends Food I know. Technol., V.13, 3-11, 2002.

RAO, Α. V .; SHIWNARAIN, N .; MAHARAJ, I. Survival of microencapsulated Bifidobacterium pseudolongum in simulated gastric and intestinal juices. Canadian Institute of Food Science and Technology Journal, v. 22, no. 4,345-49, 1989.

RIMADA, P. S .; ABRAHAM, A. G. Kefiran improves theological properties of glucono-d-lactone induced skim milk gels. International Dairy Journal, v. 16, 33-39, 2006.

ROBINSON, R.K. Survival of Lactobacillus acidophilus inferreded products-afrikanse. Suid A Frikaanse Tydskrif VirSuiwelkunde., V. 19, 25-27, 1987.

RODRIGUES, K. L .; GAUDINO CAPUTO, L. R .; TAVARESCALHO, J. C .; EVANGELIST, J .; SCHNEEDORF, J.M. Antimierobial and healing activity of kefir and kefiran extract. International Journal of Antimicrobial Agents, v. 25, 404-408, 2005.ROY1 D; GOULET, J; LEDUY, A. Continues production qf Iacticacid from whey permeate by free and calcium-alginate entrappedLactobacillus helveticus. J Dairy Sci., V. 70, 506-513, 1987.

RUAS-MADIEDO, P., R .; TUINIER, M .; KANNING, P .; ZOON.Role of exopolysaccharides produced by Lactococcus Iaetissubsp. eremoris on the viscosity of fermented milks. Int. Dairy J., v. 12, 689-695, 2002.

SALOFF-COSTE, C.J., 1996. Kefir. Danone World Newsletter, no. 11

SANDERS, M.E. Overview of functional foods: emphasis onprobiotic bacteria. Int. Dairy J., v. 8, 341-347, 1998.

SANDERS, M.E. Probiotics: considerations for human health.Nutr. Rev., v.61, no. 3, 91-99, 2003.

SANGUANSRI, P .; AUGUSTIN, M. A. Nanoseale materials development - a food industry perspective. Trends in Food Science and Technology, v. 17, no. 10, 547-556, 2006.

SCHILLINGER, U. Isolation and Identification of Iactobacilli from new probiotic type and mild yoghurts and their stability during refrigerated storage. IntJ Food Microbiol., V. 47 79-87, 1999.

SHAH, N.P .; LANKAPUTHRA, W.E.V. Improving viability of Laetobacillus aeidophilus and Bifidobaeterium ssp. in yogurt. IntDairyJ., V. 7, 349-359, 1997.SHAH1 Ν.Ρ .; LANKAPUTHRA, W.E.V; BRITZ1 M .; KYLE1 W.S.A.Survival of Lactobacillus acidophilus and Bifidobacterium Iongumin commercial yoghurt during refrigerated storage. Int Dairy J., v.5, 515-521, 1995.

SHAH, N.P .; RARULA, R.R. Microencapsulation of probioticbacteria and their survival in frozen fermented dairy desserts.Aust J Dairy Technol., V. 55: 139-144, 2000.

SHAHIDI, F .; HAN1 X. Q. Encapsulation of food ingredients. Critical Reviews in Food Science and Nutrition, v. 33, no. 6,

501-547, 1993.

SHEU, T. Y .; MARSHALL, R.T. Mieroentrapment of Iactobaeilliin calcium aginate gels. Journal of Food Science, v. 54, no. 3,557-561, 1993.

SHEU, T. Y .; Marshall, R. T .; Heymann, H. Improving survivalof culture bacteria in frozen desserts by microentrapment.Journal of Dairy Science, v. 76, no. 7, 1902-1907, 1993.

SHEU, T. Y .; Marshall, R. T .; HEYMANN, H. Improvingsurvival of culture bacteria in frozen desserts bymicroentrapment. Journal of Dairy Science, v. 76, 1902-1907,1993.SHEU1 Τ.Υ .; Marshall, R.T. Improving culture viability infused dairy desserts by microencapsulation. J Dairy Sci., V. 74.107-111.1991.

SHIMIZU, H .; MIZUGUCHI, T .; TANAKA, E .; SHIOYA, S. Nisinproduction by a mixed-system system consisting of Lactococcuslactis and Kluyveromyces marxianus. Appl. Environ. Microbiol., V. 65, no. 7, 3134/3141, 1999.

SMIDSROD, O .; SKJ AK-BRAE K, G. Alginate for cellimobilization. Trends in Food I Know Technol., V. 8, 71-75, 1990.

STEENSON, L.R .; KLAENHAMMER, T.R .; SWAISGOOD, H.E.Caleium alginate-immobilized cultures of Iaetie streptococci are protected from attack by Iytie bacteriophage. J Dairy I know., V. 70.1121-1127.1987.

SULTANA, K .; GODWARD, G .; REYNOLDS, N. Encapsulationof probiotic bacteria with alginate-starch and evaluation of survival in simulated gastrointestinal conditions and in yoghurt.International Journal of Food Mierobiology, v. 62, no. 1-2, 47-55, 2000.

SUN, W .; GRIFFITHS, M.W. Survival of bifidobacteria in yogurt and simulate gastric juice following immobilization ingellanxanthan beads. IntJ Food Mierobiol., V. 61: 17-25, 2000.SUNOHARA, H .; OHNO1 T .; SHIΒΑΤΑ, N .; SEKI1 Κ. Process for producing capsule and capsule obtained thereby. US Patent. 5: 478-570, 1995.

SURAWICZ, C.M .; ELMER, G .; SPEELMAN, P .; Van Belle, G.M.C .; FARLAND, LV .; CHINNj J. Prevention of antibiotics associated with diarrhea by Saccharomyces boulardi as prospectivestudy. Gastroenterology, v. 9, 981-988, 1989.

TANAKA, H .; MASATOSE, M .; VELEKY, I.A. Diffusioncharacteristics of substrates in calcium-alginate beads.Biotechnol Bioeng., V. 26, 53-58, 1984.

TiPAYANG, P .; KOZAKI, M. Lactic acid production by a new Lactobacillus sp., Lactobacillus vaeeinostereus Kozaki andOkada sp. nov., immobilized in calcium alginate. Journal of Fermentation Technology, v. 60, no. 6, 595-98, 1982.

TODD, R. D. Microencapsulation and flavor industry. FlavorIndustry, v. 1, no. 11, 768-771, 1970.

TRUELSTRUP-HANSEN, L .; ALLAN-WOJTAS, P.M .; JIN, Y.L.; PAULSON, AT. Survival of free and calcium-alginatemicroencapsulated Bifidobaeterium spp. in simulated gastro-intestinal conditions. Food Microbiol., V. 19, 35-45, 2002.

TUOHY, K.M .; PROBERT, H.M .; SMEJKAL, C.W .; GIBSON, G.R. Using probiotics and prebiotics to improve gut health. Drug Discovery Today, v.8, no.15, 692-700, 2003.U.S. Pat. No. 5, 175.093; 5,288,632; 5,093,253; 4,722,898; 4828,997; 5,070,019;

U.S. Pat. No. 5, 286,495; four 806.355; 4,689,293;

UBBINK, J .; Kruger, J. Physical approaches for the delivery of active ingredients in foods. Trends in Food Science and Technology, v. 17, no. 5, 244-254, 2006.

URDANETA, J .; BARRENETXE, P .; ARANGUREN, A.; IRIGOYEN, F .; MARZO, F .; IBÁNEZ, E. Intestinal benefits from kefir-supplemented diet in rats. Nutrition Research, v. 27, no. 10, 653-658, 2007.

VINDEROLA, C.G .; BAILO, N .; REINHEIMER, J.A. Survival of probiotic microflora in Argentinian yoghurts during refrigerated storage. Food Res Int., V. 33: 97-102, 2002.

VINDEROLA, C.G .; DUARTE, J .; THANGAVEL, D .; PERDIGÓN, G .; FARNWORTH, E .; MATAR, C. Immunomodulating capacityof kefir. J. Dairy Res., V. 72, 195-202, 2005.

VINDEROLA, G .; DANGER 'N, G .; DUARTE, J .; FARNWORTH, E .; MATAR, C. Effects of the oral administration of theexopolysaccharide produced by Lactobacillus kefiranofaciens onthe gut mucosal immunity. Cytokine, v. 36, 254-260, 2006. WENRONG, S .; GRIFFITHS, M.W. Survival of bifidobacteria inyogurt and simulated gastric juice following immobilization ingellan-xanthan beads. IntJ Food Microbiol., V.61: 17-26, 2000.

YOO, I.K .; SEONG, G. H .; CHANG1 H. N .; PARK, J. K. Encapsulation of Lactobacillus casei cells in liquid-core alginatescapsules for Iactic aeid produetion. Enzyme and MicrobialTechnology, v. 19, no. 6, 428-33, 1996.

ZHOU, Y .; MARTINS, E .; GROBOILLOOT, A .; CHAMPAGNE, C.P; NEUFELD, R.J. Spectrophotometric quantification of Iaetieaeid bacteria in alginate and control of cell release with chitosancoating. J Appl Microbiol., V. 84, 342-348, 1998.

ZIEMER, C.J .; GIBSON, G.R. An overview of probiotics, prebiotics and synbiotics in the functional food concept: perspectives and future strategies. Int. Dairy J., v. 8, 473-479,1998.

ZUBILAGA, M .; WEILL, R .; POSTAIRE, E .; GOLDMAN, C.; CARO, R .; BOCCIO, J. Effect of probiotics and functional foods and their use in different diseases. Nourish Res., V. 21, n. 3,569-579, 2001.

Claims (7)

1. "INDUSTRIAL PROCESS OF! MOBILIZATION OF A MICRORGANISM CONTAINMENT PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES, THROUGH MICROCAPLES ALGINATE present in the microcapsules forming microcapsules and forming microcapsules in their microorganisms. allow the survival of microorganisms, does not depend on the constant maintenance characteristic of kefir and can be used to make probiotics food, cosmetics and medicines without changing taste, odor or color. The process comprises the following steps: a) Cultivation of a microorganism pool on different substratessoya extract, grape juice, lemon and plum juice, deabacaxi juice, tangerine juice, passion fruit juice, acai juice, apple juice, juice pear juice, peach juice, coconut water, dean juice; (b) After this, the culture is added to a solution formed by sodium alginate, and / or Kefiran and / or Pectin forming a new solution (solution A). This solution is kept under stirring until use (1); c) At the time of use (2), solution A is transferred to tank 2 where it is aspirated and dosed by a peristaltic pump, which drips it into a second solution (solution B), the latter of calcium chloride; (d) Once a certain amount of microcapsules has been produced (3), which may be either time or volume, this tank shall be replaced by tank III / 3, which shall contain only calcium chloride solution and shall receive the dripping. the peristaltic pump; (e) The microcapsules of tank 111/1, which has been replaced, shall be allowed to stand for 5 to 20 minutes, which are necessary to supplement the pelletisation process. f) After this time of rest, the pellets and the solution will be poured into a sieve, where the liquid and solid phases will be separated, leaving the liquid phase in tank III / 2, which will replace the peristaltic pump drop tank (4). ) and repeat the process for the production of more pellets; g) On the other hand, the collected pellets will be taken to tank IV (5), where, without being removed from the sieve, they will be properly washed with distilled water in a closed circuit, concluding with this operation a cycle of the production of kefir pellets. 2. "INDUSTRIAL PROCESS OF! MOBILIZING A MICRORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES BY MEASURING CALCIUM ALGINATE ALGINATEES", according to claim 1 and claim 1 . 3. "INDUSTRIAL PROCESS OF! MOBILIZATION OF A MICRORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES BY MEASURING CALCIUM MODIFIED ALGINATE", according to claim 1, according to claim 1. yeast 4. "INDUSTRIAL PROCESS OF! MOBILIZATION OF A MICRORGANISM COUNCIL PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES, THROUGH CALCULOMOMIFIED ALGINATE ALUMINATION", according to claim 1, characterized by the homogeneous product 1 Lactobacilli: Lactobacillusbrevis; Lactobacilluscellobiosis; Lactobacillus aeidophillus; Lactobacillus casei ssp. laetosus; Lactobacillus casei SSP.rhamnosus; Laetobacillus casei; Lactobacillus paracasei SSP.paraeasei; Lactobacillus helveticus SSP. lactis; Lactobacillusdelbrueckii SSP. lactis; Lactobacillus delbrueckii SSP. bulgaricus; Lactobaciilus lactis; Lactobacillus fructivorans; Lactobacillus rilgardii; Lactobacillus kefir; Lactobacillus kefiranofaciens; Lactobacilluskefirgranum SP. ; Lactobacillus parakefir SP; 2-Lactococcus / Streptococcus: Lc. lactis ssp. lactis; Lc lactis var.diacetylactis; Lc lactis ssp. cremoris; S. sallivarius ssp.thermophilus; S. lactis; Enterococcus durans; Leuconostoc cremoris; L. mesenteroids; 3- Yeast: Kluyveromyces lactis; Kluyveromyces marxianus var. Marxianus; K. bulgaricus; K. fragilis Candida kefir; C. pseudotropicalis; C. rancens; Saccharomyceslactis; A- Acetobacter: Acetobacters aceti; A. rasens. 5.) "INDUSTRIAL PROCESS OF! MOBILIZATION OF A MICRORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES, BY MEASURING CALCIUM ALGINATE ALGINATED, FACTORY OF A MIXTURE OF Sodium alginate kefiranoe or pectin applied in the method of claim 1. 6.) "INDUSTRIAL PROCESS OF! MOBILIZING A MICROORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS, AS WELL AS ITS BIOACTIVES BY MEASURING CALCIUM ALGINATE MICROCAPULES", according to claim 1, as produced by the microcapsulated method of claim 1, which is produced by the method 1. at temperatures in the range of 25 to 50 ° C. 7.) "INDUSTRIAL PROCESS OF! MOBILIZING A MICROORGANISM CONSERVATION PRESENT IN THE KEFIRBIOLOGICUS AS WELL AS ITS BIOACTIVES BY MEASURING CALCIUM MODIFIED ALGINATED MICROCAPSULES PROCEDURE BY THE MODELING OF THE CALCULOMODIFIED MEASUREMENT 1 -5 ° C and then dried in greenhouses at temperatures in the range 25 to 50 ° C.