MXPA99011241A - Carbohydrate-based enzyme granulates - Google Patents

Abstract

A process for the preparation of an enzyme-containing granulate is disclosed where an aqueous enzyme-containing liquid is mixed with an edible carbohydrate-based solid carrier, such as starch, mechanically processed into granules, and subsequently dried. This enzyme granulate is suitable for the manufacture of animal feed compositions by mixing feed ingredients with the granulate, treating with steam and pelleting. The compositions show improved enzyme stability during the pelleting process.

Description

ENZYMATIC GRANULES BASED ON CARBOHYDRATE FIELD OF THE INVENTION The present invention relates to the formulation of enzymes, preferably food enzymes, in granules containing carbohydrate (for example starch), and to processes for the preparation of such enzyme-containing granules. These granules (edible) can then be used in animal feeds.

BACKGROUND OF THE INVENTION The use of various enzymes in the feeding of animals, for example cattle, has become almost common practice. These enzymes are usually produced by the cultivation of microorganisms in large-scale fermenters operated by industrial producers of enzymes. At the end of the fermentation, the resulting "broth" is usually subjected to a series of filtration steps to separate the biomass (microorganisms) from the desired enzyme (in solution) The enzyme solution is either sold as a liquid REF .: 32198 (often after the addition of various stabilizers) or processed to form a dry formulation. The liquid and anhydrous enzyme formulations are used on a commercial scale by the food industry. The liquid formulations can be added to the food after becoming pellets, in order to avoid heat inactivation of the enzyme (s), which could occur during the pellet formation process. However, the amounts of enzyme in the final food preparations are usually very small, which makes it difficult to achieve a homogeneous distribution of the enzyme in the food, and the liquids are notoriously more difficult to mix uniformly than the anhydrous ingredients. In addition, specialized (expensive) equipment is needed to add liquids to the feed after pellet formation, which is not currently available in most food mills (due to the extra cost). Anhydrous or dried enzyme (s) formulations, on the other hand, have the disadvantage of heat inactivation of the enzymes during the pellet formation. Preferred manufacturing protocols in the food industry involve the formation of pellets by steam, where the food is subject to injection or steam injections prior to the formation of the pellets. In the subsequent formation of the pellets, the feed is forced through a die or die and the resulting strips are cut into suitable pellets of variable length. HE. Moisture content immediately before the formation of the pellets is generally between 18% and 19%. During this process temperatures can rise to 60-95 ° C. The combined effect of high moisture content and high temperature is harmful to most enzymes. These disadvantages are also found in other types of thermomechanical treatments such as extrusion and expansion. In order to address and overcome these problems European Patent EP-A-0, 257, 996 (Cultor Ltd) suggests that the stability of enzymes in food processing could be increased by the preparation of an enzyme 'premix' where an enzyme-containing solution is absorbed onto a grain-based carrier consisting of flour, and the premix is subsequently pelletized and dried, however, these flour-based premixes are not suitable for milder processing methods (from the premix). similar to dough) in granulates, such as low pressure extrusion or high cut granulation, due to the gummy or sticky character of the flour-based premixes Several enzyme manufacturers have developed alternative formulation methods to try to improve stability of dry or anhydrous enzyme products during pellet formation and storage European Patent EP-A-0 569, 468 (Nov. or Nordis) refers to a formulation consisting of a "T-granulate" containing enzyme that is coated with a high melting point wax or a fat that is said to improve resistance to pelletizing conditions. The granulate is prepared by mixing an anhydrous inorganic filler (for example sodium sulfate) with the enzyme solution in a high cut granulator. The "European Patent EP-A-0, 569, 468 teaches that any beneficial effect of the coating with respect to the stability of the pellet formation is specific to the type of coated granulate, which in this case is based on a sodium sulphate filler. However, the absorption capacity of these fillers (sodium sulfate) is much lower than that of the carriers such as flour, which is undesirable if someone wishes to produce granules containing enzyme, more concentrates. In addition, the granulates have a broad particle size distribution, which makes it difficult to obtain a homogenous enzyme concentration at all lengths. In addition, the bioavailability of the enzyme to the animal is diminished by the coating with wax or fat. WO-A-97/16076 (Novo Nordisk) also refers to the use of waxes and other particulate water insoluble substances, but here they are used as a matrix material. There is thus a need for stable formulations of enzymes that are based on a carrier that is suitable for granulation methods other than pelletization, and which may have a high absorption capacity.

DESCRIPTION OF THE INVENTION In a first aspect of the present invention, there is provided a process for the preparation of an enzyme-containing granulate, suitable for use in an animal feed, the process comprising, the processing of an enzyme, a solid carrier comprising at least 15% (w / w) of an edible carbohydrate polymer and water "" in appropriate relative amounts to obtain granules containing enzyme, and subsequently drying the granules. The granulate containing enzyme produtible by this process (which forms the second aspect of the invention, which also covers a granulate comprising dry granules formed from an enzyme and a solid carrier which comprises at least 15% (p. / p) of an edible carbohydrate polymer) seeks to solve or at least mitigate the problems encountered in the prior art. - The invention can thus provide processes for the preparation of enzymatic formulations in the form of granules using the granulate as a carrier. The carrier can be in particulate or powder form. The enzyme and water are preferably provided as an enzyme-containing liquid (preferably aqueous), such as a solution or suspension, which can be mixed with the solid carrier and allowed to absorb onto the carrier. During or after mixing, the liquid containing the enzyme and the carrier are processed to form a granulate, which can then be subsequently dried. The use of the carbohydrate carrier can allow the absorption of large amounts of enzyme-containing liquid (and therefore the enzyme). The mixture can be used to form a plastic paste or a non-elastic mass that can be easily processed into granules, for example it is extrudable. Suitably, the carrier is non-fibrous, which allows for easier granulation: fibrous materials can prevent extrusion granulation. A number of prior art documents refer to pellets containing various enzymes, but they find use as detergents, often in washing compositions. In contrast, the present application finds use in animal feeds and for that reason the granulates of the invention are edible (by animals) and preferably also digestible. It will therefore not be surprising if the granulates, granules and compositions of the invention are free of soap, detergents and bleaching agents or bleaching compounds, zeolites, binders, fillers (Ti02, kaolin, silicates, talc, etc.). ) to name just a few. The edible carbohydrate polymer must be chosen such that it is edible by the animal for whom the food is intended, and preferably also digestible. The polymer preferably comprises glucose (for example, a polymer containing glucose), or units (CeHioOs). Preferably, the carbohydrate polymer comprises units of α-D-glucopyranose, amylose (a polymer of linear (1->) -aD-glucan) and / or amylopectin (a branched D-glucan with links to D- (1-4) and aD- (l- »6)). Starch is the preferred carbohydrate polymer. Other suitable glucose-containing polymers, which may be used in place of, or in addition to, starch, include α-glucans, β-glucans, pectin (such as proto-pectin), and glycogen. Derivatives of these carbohydrate polymers, such as ethers and / or esters thereof, are also contemplated although the gelatinized starch is avoided as much as possible and thus can not be present. Suitably, the carbohydrate polymer is insoluble in water. Corn, potato and rice starch are used in the examples described herein. However, starch obtained from other sources (for example, plants such as vegetables or crops), such as tapioca, cassava, wheat, corn, sago, rye, oats, barley, yam, sorghum, or maranta is equally applicable. Similarly, native or modified starch types (e.g., dextrin) can be used in the invention. Preferably, the carbohydrate (eg, starch) contains little or no protein, for example less than 5% (w / w), such as less than 2% (w / w), preferably less than 1% (w / w). Notwithstanding the type of starch (or other carbohydrate polymer), it must be in a form that allows it to be used in an animal feed, in other words an edible or digestible form. At least 15% (w / w) of the solid carrier can comprise the carbohydrate polymer (such as starch). Preferably, however, at least 30% (w / w) of the solid carrier comprises the carbohydrate, optimally at least 40% (w / w). Advantageously, the major component of the solid carrier is the carbohydrate (for example starch), for example more than 50% (w / w), preferably at least 60% (w / w), suitably at least 70% (w / w) , and optimally at least 80% (w / w). These percentages by weight are based on the total weight of the non-enzymatic components in the final dry granulate. In the process of the present invention, the enzyme and water may be present in the same composition before being contacted with the solid carrier. In this aspect, an aqueous liquid containing enzyme can be provided. This liquid may be a solution or a suspension that is derived from or derived from, a fermentation process. This fermentation process will usually be one in which the enzyme is produced. The fermentation process can result in a broth containing the microorganisms (which produce the desired enzyme) and an aqueous solution. This aqueous solution, once separated from the microorganisms (for example, by filtration) can be the aqueous liquid containing enzyme used in the invention. Thus, in the preferred embodiments the aqueous liquid containing enzyme is a filtrate.

The amount of enzyme-containing liquid (and thus the enzyme) that can be absorbed onto the carrier is usually limited by the amount of water that can be absorbed. For natural, granular starch, this can vary between 25 and 30% (p / p), without using elevated temperatures (which promote starch swelling). In practice, the percentage of enzyme fluid that is going to be added to the carbohydrate will often be much greater than this, because the enzyme-containing liquid will usually contain a significant amount of solids. The enzyme solution may contain about 25% (w / w) solids, as a result of which the carbohydrate (eg, starch) and the enzyme solution can be mixed at a carbohydrate: enzyme solution ratio of 0.5: 1 to 2. : 1, for example 1.2: 1 to 1.6: 1, such as at a ratio of approximately 60% (w / w): 40% (w / w), respectively. Preferably, the amount of liquid added to the solid carrier is such that __ (substantially) all the water in the liquid (aqueous) is absorbed by the carbohydrate present in the solid carrier. At elevated temperatures, starch and other carbohydrate polymers can absorb much greater amounts of water under swelling. For this reason the carbohydrate polymer is desirably capable of absorbing water (or aqueous liquids containing enzyme). For example, corn starch can absorb up to three times its weight of water at 60 ° C and up to ten times at 70 ° C. The use of higher temperatures in order to absorb a larger amount of enzyme-containing liquid is thus contemplated by the present invention, and of course it is especially preferable when it is associated with thermostable enzymes. For these enzymes, therefore, the mixture of the solid carrier and the liquid (or enzyme and water) can be conducted at elevated temperatures (for example above room temperature), such as above 30 ° C, preferably above 40 ° C and optimally above 50 ° C. Alternatively, or in addition, the liquid can be provided at this temperature. However, in general, non-swelling conditions are preferred at lower temperatures (for example environment). This can minimize the loss of activity that arises from the instability of enzymes (sensitive to heat) at higher temperatures. Suitably the temperature during the mixing of the enzyme and water is 20 to 25 ° C.

The mechanical processing used in the present invention for the preparation of the mixture of the enzyme, water (for example an enzyme-containing liquid) and the solid carrier in granules (in other words, granulation) can employ known techniqfrequently used in processes of food, food and enzyme formulation. This may comprise expansion, extrusion, spheronization, pelletization, high-cut granulation, drum pelletization, fluid bed agglomeration or a combination thereof. These processes are usually characterized by a mechanical energy input, such as the drive of a screw, the rotation of a mixing mechanism, the pressure of a rotating mechanism of a pelletizing apparatus, the movement of particles by a plate of rotating bottom of a fluidized bed agglomerator or movement of the particles by a gaseous stream, or a combination thereof. These processes allow the solid carrier (for example in the form of a powder) to be mixed with the enzyme and with water, for example a liquid containing enzyme (an aqueous solution or suspension), and subsequently granulated.

Alternatively, the solid carrier can be mixed with the enzyme (for example in a powder form) to which water is then added, such as a liquid (or suspension) (which can act as a granulation liquid). In a further embodiment of the invention the granulate (for example an agglomerate) is formed by spraying or coating enzyme containing liquid on the carrier, such as in a fluidized bed agglomerator. Here, the resulting granules may include an agglomerate as may be produced in a fluidized bed agglomerator. Preferably, the mixture of the enzyme-containing liquid and the solid carrier further comprises mixing the mixture.This can improve the plasticity of the mixture in order to facilitate granulation (for example extrusion) .If the granulate is formed by extrusion This is preferably done at low pressure This may offer the advantage that the temperature of the mixture that is extruded will not increase, or will only increase slightly Extrusion at low pressure includes extrusion for example in a basket extruder or Fuji Paudal dome Preferably, extrusion does not result in the temperature of the material being extruded rising above 40 ° C. Extrusion can naturally produce granules (the granules can break after passing through a matrix or die) or a cutter may be employed, suitably, the granules will have a water content of 30 to 40%, such as 33 to 37%. it is preferably from 3 to 15, such as from 5 to 12% (for example at least 50,000 ppm). The granules obtained can be subjected to rounding (for example spheronization), such as in a spheronizer, for example, a machine MARUMERISERMR and / or compaction. The granules can be spheronized before drying, since this can reduce the formation of dusts in the final granulate and / or can facilitate any coating of the granulate. The granules can then be dried, such as in a fluidized bed dryer or, in the case of fluidized bed agglomeration, they can be immediately dried (in the agglomerator) to obtain granulates (dry, solid). Other known methods for the drying of granules in the food, food or enzymes industry can be used by the skilled person. Suitably, the granulate is capable of flowing. Drying preferably takes place at a temperature of 25 to 60 ° C, such as 30 to 50 ° C. Here drying can last from 10 minutes to several hours, such as 15 to 30 minutes. The length of time required will of course depend on the amount of granules that are to be dried, but as a guide this is from 1"" to 2 seconds per kg of granules. After drying the granules, the resulting granulate preferably has a water content of 3 to 10%, such as 5 to 9%. A coating may be applied to the granulate to give additional characteristics or properties (eg flavored), such as low dust content, color, surrounding enzyme protection, different enzyme activities in a granulate or a combination thereof. The granulates can be coated with a fat, wax, polymer, salt, ointment and / or a coating (for example liquid) containing an enzyme (second) or a combination thereof. It will be apparent that if desired, several layers of (different) coatings can be applied. To apply the coating or coatings on the granulates a number of known methods are available, which include the use of a fluidized bed, a high cut granulator, a mixer granulator, or a Nauta mixer. In other additional embodiments, the ingredients may be incorporated into the granulate, for example as processing aids, for further improvement of the pelletizing stability and / or storage stability of the granulate. A number of such preferred additives are discussed below. Salts may be included in the granulate (for example, with the solid carrier or water). Preferably (as suggested in European Patent EP-A-0 758, 018) one or more inorganic salts can be added, which can improve the stability in the processing and storage of the anhydrous enzyme preparation. The preferred inorganic salts are soluble in water. These may comprise a divalent cation, such as zinc (in particular), magnesium, and calcium. Sulfate is the most favored anion, although other anions can be used which result in water solubility. The salts (for example to the mixture) in the solid form can be added. However, the salt (s) can be dissolved in the water or in the liquid containing the enzyme, before mixing with the solid carrier. Suitably, the salt is provided at an amount that is at least 15% (w / w based on the enzyme), such as at least 30%. However, this can be as high as at least 60% or even 70% (again, w / w based on the enzyme). These quantities can be applied either to the granules or to the granulate. The granulate can therefore comprise less than 12% (w / w) of the salt, for example from 2.5 to 7.5%, for example from 4 to 6%. If the salt is provided in the water, then it may be in an amount of 5 to 30% (w / w), such as 15 to 25%. Further improvement of the stability of the pellet formation can be obtained by the incorporation of hydrophobic, gel forming or slow dissolving compounds (for example in water). These may be provided from 1 to 10%, such as from 2 to 8%, and preferably from 4 to 6% by weight (based on the weight of the water and the solid carrier ingredients.) Suitable substances include derivatized celluloses. , such as HPMC (hydroxyprvyl methylcellulose), CMC (carboxymethylcellulose), HEC (hydroxyethylcellulose); polyvinyl alcohols (PVA); and / or edible oils. Edible oils, such as soybean oil or cane oil, can be added (for example to the mixture to be granulated) as a processing aid, although it will often be preferred that the granulate does not contain any hydrophobic substance ( for example, palm oil). Preferably, the granules have a relatively narrow size distribution (for example these are monodispersed). This can facilitate a homogenous distribution of the enzyme in the granules and / or in the enzyme granulate in the animal feed. The process of the invention tends to produce granules with a narrow size distribution. However, if necessary, an additional step in the process can be included to further narrow the size distribution of the granules, such as sieving. The size distribution of the granulate is suitably between 100 μm and 2000 μm, preferably between 200 μm and 1800 μm, and optimally between 300 μm and 1600 μm. The granules may be irregular (but preferably regular), for example approximately spherical. The water or the liquid containing the enzyme may comprise one or more enzymes, and are usually of microbial origin, for example, obtained from a microbial fermentation. Usually, the enzyme will be in an active form (for example this may have catalytic or physiological activity). Preferably the liquid is in a concentrated form, such as an ultrafiltrate (UF), which may allow the production of a granulate with a desired level of activity. The appropriate enzyme (s) are those that are to be included in the animal feed that includes pet food. The function of these enzymes is often to improve the feed conversion rate, for example, by reducing the viscosity or by reducing the antinutritional effect of certain food compounds. Food enzymes (such as phytase) can also be used, such as to reduce the amounts of compounds that are "harmful to the environment in the manure." Preferred enzymes for these purposes are: phosphatases, such as phytases (3-phytases and phytases) and / or acid phosphatases, carbohydratases, such as amylolytic enzymes and plant cell wall degrading enzymes, of which are included cellulases such as β-glucanases, hemicellulases such as xylanases, or galactanases; peptidases, galactosidases, pectinases, esterases, proteases, preferably with an optimum neutral and / or acid pH, and lipases, preferably phospholipases such as mammalian pancreatic phospholipases A. Preferably, the enzyme does not include starch degrading enzymes (for example amylases). Proteases may be excluded because they can cause damage if they are ingested, if the enzyme is a phosphatase, such as a na phytase, then preferably the final granulate will have an activity of 5,000 to 10,000 such as 6,000 to 8,000, FTU / g. If the enzyme is a plant cell wall degrading enzyme, for example a cellulase, and in particular a hemicellulase such as xylanase, then the final granulate can have an enzyme activity in the range of 3,000 to 100,000, preferably 5,000 to 80,000, and optimally from 8,000 to 70,000, EXU / g. If the enzyme is a cellulase, such as β-glucanase, then the final granulate can have an enzymatic activity of 500 to 15,000, preferably 1,000 to 10,000, and optimally of 1,500 to 7,000, BGU / g. The granulates may comprise from 5 to 20, for example from 7 to 15% of the enzyme (s). The enzyme (s) can be of natural or recombinant origin. In addition to these enzymes the invention is equally applicable to polypeptides with other biological activities, such as antigenic determinants, for example that find use in vaccines and / or genetically engineered polypeptides having an increased content of essential amino acids, of which biological activity may be sensitive to thermal inactivation, and the term "enzyme" as used herein has to be considered accordingly.A preferred process according to the invention therefore comprises; a) the mixture of water, enzyme and solid carrier comprising at least 15% (w / w) or an edible carbohydrate polymer, for example the mixture of the solid carrier with an aqueous, enzyme-containing liquid; b) the optional mixing of the resulting mixture; c) granulation, for example by mechanical processing of the mixture, in order to obtain granules containing enzyme, for example by the use of a granulator or by extrusion; d) the optional spheronization of the granules; e) drying the resulting granules to obtain a granulate containing enzyme. During the complete process, attention can be directed to maintaining the maximum temperature at which or the enzymes are exposed to less than 80 ° C. The granulates of the invention are suitable for use in the preparation of an animal feed. In such processes the granulates are mixed with food substances, either as such, or as part of a premix. The characteristics of the granulates according to the invention allow their use as a component of a mixture which is very suitable as an animal feed, especially if the mixture is treated by steam and subsequently converted into pellets. The dry granules may be visible or distinguishable in such pellets. Thus, a third aspect of the present invention relates to a process for the preparation of animal feed, or a premix or a precursor for an animal feed, the process comprises mixing a granulate of the second aspect, with a or more food substances for animals (for example seeds) or food ingredients. This can then be sterilized, for example, subject to heat treatment. The resulting composition is then suitably processed in the form of pellets. A fourth aspect of the invention relates to a composition comprising a granulate of the second aspect, which composition is preferably an edible food composition such as an animal feed. This composition is preferably in the form of pellets (there may be 1-5, for example 2-4 dry pellets per pellet). The composition may have a water content of 10 to 20%, for example 12 to 15%. The amount of enzyme or enzymes is suitably from 0.0005 to 0.0012%, such as at least 5 ppm.

A fifth aspect relates to a process for promoting the development of an animal, the process comprising feeding an animal with a diet comprising a granulate of the second aspect, or a composition of the fourth aspect. Here, the animal diet can include either the granulate itself, or the granulate present in a food. Suitably, the composition comprises from 0.05 to 2.0, such as from 0.3 to 1.0, optimally from 0.4 to 0.6 FTU / g of a phosphatase, for example a phytase. A xylanase from 0.5 to 50, for example from 1 to 40 EXU / g, may be present. Alternatively or in addition, a cellulase of 0.1 to 1.0 may be present, for example 0.2 to 0.4 BGU / g. A sixth aspect of the present invention relates to the use of the granulate of the second aspect in, or eats a component of, an animal feed or for use in an animal diet. A seventh aspect of the present invention relates to the use of a composition comprising at least 15% (w / w) of an edible carbohydrate polymer as a carrier for an enzyme, to improve the stability of pellet formation, the enzyme.

Suitable animals include farm animals (pigs, poultry, livestock), non-ruminant or monogastric animals (pigs, poultry, marine animals such as fish), ruminants (cattle or sheep, for example cows, sheep, goats, deer, calves, rams). Poultry includes chickens, chickens and turkeys. The preferred features and characteristics of one aspect of the invention are equally applicable to others, muta ti s mu tandi s. The following examples are presented merely to illustrate the invention, and are not intended to be considered as limiting.

EXAMPLES Materials and General Methods Extrusion tests were performed using a Fuji Paudal DG-Ll basket extruder, with 1.0 mm mesh openings, 1.2 mm mesh thickness, with an operating speed of 70 rpm, and a current of 0.6-2.0 A. The spheronizer It was a Fuji Paudal Marumerizer QJ-400, with a 3-liter cargo volume, 3 mm plate opening, 45-200 seconds retention time and 750 rpm rotation speed. _ The high cut granulation tests were conducted using a Lodige type FM20 high cut granulator, with a shredder speed of 1500 rpm and a plowing speed of 100 rpm. Powder was placed in the granulator and the enzyme-containing liquid was sprayed on top. The resulting granules were dried in a fluidized bed dryer. The enzymatic solutions used were: an ultrafiltrate of a phytase derived from Apergi l l us with an activity of 16840 FTU / g, and a dry solids content of 22.4% (w / w) (Examples 1 to 7). _ an ultrafiltrate containing a mixture derived from Tri choderma of endo-xylanase and β-glucanase with activities of 12680 EXU / g and BGU / g, and a dry solids content of 20.6% (w / w) (Example 8). The activity of phytase was determined according to the procedure 'ISL-Method 61696' (manual vanadate test) The activity of β-glucanase was determined according to the procedure 'ISL-Method 62170' (manual viscosimetric assay). The activity of endo-xylanase was determined according to the procedure 'ISL-method 62169' (manual viscosimetric assay) .The ISL methods are obtainable at the request of Gist-brocades, Food Specialties, Agri Ingredients Group, agesse eg 1, PO Box 1 , 2600 MA, Delft, Holland.

EXAMPLE 1 Preparation of enzyme granules based on corn starch, by kneading, extrusion, feronization and drying An enzyme preparation was obtained by mixing and kneading a mixture of 60% (w / w) corn starch with 40% (w / w) of an ultrafiltrate containing phytase. This mixture was extruded using the Fuji Paudal basket extruder to obtain a wet extrudate which was then spheronized in the MARUMERISERMR for one minute to obtain round particles with an average diameter of 780 μm. These particles were subsequently dried in a fluid bed dryer for 20 minutes at a bed temperature of 40 ° C, and an inlet temperature of 75 ° C. About 500 kg of the granules were dried in 18 minutes. The dry enzyme granulate obtained in this way had an activity of 6980 FTU / g.

EXAMPLE 2 Preparation of an enzymatic granulate based on corn starch, by high-cut granulation and drying The ultrafiltrate containing phytase and corn starch were blended in a high cut type granulator in Lddige type batches, with a batch size of 20 liters. The granulator was filled with 60% (w / w) of corn starch and 40% (w / w) ultrafiltrate, and was sprayed into the mixer during the mixing process. After the addition of the ultrafiltrate (10 minutes) the granulator continued mixing for another 5 minutes to allow the particles to be formed and compacted. The granules obtained in this way were dried in a fluidized bed dryer as in Example 1. The resulting granulate had an activity of 7420 FTU / g. The average diameter of the particles was 480 μm.

EXAMPLE 3 Preparation of an enzyme granule based on corn starch by mixing, pelletizing and drying - A mixture of 40% (w / w) of phytase ultrafiltrate and 60% < p / p) of corn starch. "The mixture was converted into pellets using a Schlütter press type PP85, where the extrudates were cut by rotary blades in the extruder head, with a die plate containing holes 1 mm in diameter. The pellets were dried as in Example 1, resulting in a final product with an activity of 7460 FTU / g.The mean diameter of the particles was 1080 μm.

EXAMPLE 4 Preparation of an enzyme granulate based on potato starch containing soybean oil and additions of MgSO4 by mixing, kneading, pelletizing and drying In a mixer / kneader 30 kg of potato starch and 2.5 kg of soybean oil were added and mixed. Subsequently the phytase ultrafiltrate was added containing MgSO4 * 7H20 (3.5 kg of MgSO4 # 7H20 were dissolved in 14 kg of ultrafiltrate). The product was mixed thoroughly in the kneader, then extruded and dried in a fluid bed dryer as in Example 1. This resulted in a product of 5870 FTU / g.

EXAMPLE 5 Preparation of an enzyme granulate based on rice starch by mixing, kneading, extruding, spheronizing and drying A mixture was prepared by mixing and kneading 62% (w / w) of rice starch and 38% (w / w) of the phytase ultrafiltrate. This mixture was extruded using the Fuji Paudal basket extruder to obtain a wet extrudate which was then spheronized in the MARUMERISERMR for one minute to obtain round particles with an average diameter of 785 μm. These particles were subsequently dried in a fluidized bed dryer as in Example 1. The final activity of the granulate was 7280 FTU / g.

EXAMPLE 6 Preparation of an enzyme granulate based on corn starch containing "an addition of HPMC by mixing, kneading, extrusion, feronization and drying An enzyme preparation was obtained by kneading a mixture of 54% (w / w) of corn starch, 5% HPMC (hydroxypropylmethylcellulose) and 41% (w / w) of a phytase ultrafiltrate. - This mixture was extruded using the Fuji Paudal basket extruder to obtain a wet extrudate which was spheronized in the MARUMERISERMR for one minute, to obtain round particles of an average diameter of 780 μm, which were subsequently dried in a bed dryer. fluidized for 20 minutes at a bed temperature of 40 ° C, and inlet temperature of 75 ° C. The dried enzyme granulate obtained in this way had an activity of 8470 FTU / g.

EXAMPLE 7 Preparation of an enzyme granulate based on corn starch containing an HEC addition by mixing, kneading, extrusion, spheronization and drying An enzyme preparation was obtained by mixing and kneading 54% (w / w) of corn starch, 5% (w / w) of HEC (hydroxyethylcellulose) with 41% (w / w) of the phytase ultrafiltrate. This mixture was extruded using the Fuji Paudal basket extruder to obtain a wet extrudate which was spheronized in the MARUMERISERMR for one minute to obtain round particles with an average diameter of 780 μm. These were subsequently dried in a fluid bed dryer for 20 minutes at 40 ° C bed temperature, and inlet temperature of 75 ° C. The dry enzyme granulate obtained in this way had an activity of 8410 FTU / g.

EXAMPLE 8 Preparation of an enzyme granulate based on corn starch by high-cut granulation and drying In a high-cut granulator in batches of the Lódige type, with a batch size of 20 liters, 60% (w / w) of corn starch was mixed with 40% (w / w) of the ultrafiltrate containing endo-xylanase and β-glucanase, as follows. The granulator was filled with the corn starch and the ultrafiltrate was sprayed on the mixer during the mixing process. After the addition of the ultrafiltrate (10 minutes) the granulator was operated for another 5 minutes to allow the particles to be formed and compacted. The granules obtained in this way were dried in a fluidized bed dryer as in Example 1. The resulting granulate had an activity of 13100 EXU / g and 5360 BGU / g.

EXAMPLE 9 Comparison of the stabilities of the formation of pellets The different enzyme granulates of the invention were subjected to a pellet formation test and their stability in pellet formation was compared with those of the standard food enzyme formulations. The "pellet" test consisted of mixing the enzyme (granulate) with a feed premix at 1000 ppm This mixture was treated by steam injection to give a temperature rise at 70 ° C, after which the mixture It was converted into pellets in a pellet forming machine to obtain the food pellets, which were subsequently dried. "This type of process is typical for the food industry, to obtain pellets. For NATUPH0SMR, a formulation containing phytase , used as a standard, for comparison, was a mixture of wheat bran with spray-dried ultrafiltrate.

For NATUGRAINMR, an enzyme preparation containing ß-glucanase and endo-xylanase, this is a granule prepared by fluidized bed, prepared by coating a saline core with an enzyme layer, applied by spraying the core with an ultrafiltrate. Table 1 summarizes the results of the pellet formation tests. It is apparent from Table 1 that granulates made using a carbohydrate carrier gave improved pelletizing yields when compared with standard formulations.

Table 1: Results of the pellet formation tests It is clear from "Table 1 that the type of granulation method, for example mechanical processing, is not critical to face the problems to be solved by the invention." The formulations using the carbohydrate polymer provided a stability in the formation of pellets much better, compared to the known formulations of NATUPHOSMR and NATUGRAINMR.

It is noted that in relation to this date, the best method known to the applicant for carrying out the aforementioned invention is that which is clear from the present description of the invention.

Claims (27)

RE IVINDICATIONS Having described the invention as above, the content of the following claims is claimed as property:
1. - 1. A process for the preparation of a granulate containing enzyme, suitable for use in a -food for animals, the process is characterized in that it comprises the processing of an enzyme, a solid carrier comprising at least 15% (p / p) of an edible carbohydrate polymer and water, in appropriate relative amounts to obtain granules containing "enzyme, and subsequently drying the granules.
2. 2. A process according to claim 1, characterized in that the water and the enzyme are provided as an aqueous liquid containing enzyme.
3. 3. A process according to claim 2, characterized in that the liquid is a filtrate derived from a fermentation process that results in the production of the enzyme.
4. 4. A process according to any of the preceding claims, characterized in that the process comprises: a) the mixture of an aqueous liquid containing the enzyme, with the solid carrier; b) the mechanical processing of the mixture obtained in a) to obtain granules containing enzyme; and _ c) drying the enzyme-containing granules obtained in step (b).
5. 5. A process according to any of the preceding claims, characterized in that the mechanical processing and comprises extrusion, pelletization, high-cut granulation, expansion, fluidized-bed agglomeration or a combination thereof.
6. 6. A process according to any of the preceding claims, characterized in that an aqueous liquid containing enzyme and the solid carrier are mixed and the resulting mixture is kneaded before granulation.
7. 7. A process according to any of the preceding claims, characterized in that the processing is extrusion carried out at low pressure and / or in a basket or dome extruder.
8. 8. A process according to any of the preceding claims, characterized in that the granules obtained are spheronized before drying.
9. 9. A granulate containing enzyme, produced by a process as defined according to any of the preceding claims.
10. 10. A granulate comprising dry granules, characterized in that they are formed from an enzyme and a solid carrier which comprises at least 15% (w / w) of an edible carbohydrate polymer.
11. 11. A granulate according to claims 9 or 10, "characterized in that the granules comprise at least one divalent cation.
12. 12. A granulate according to any of claims 9 to 11, characterized in that the granules comprise one or more hydrophobic compounds, gel formers or water insoluble.
13. 13. A granulate according to claim 12, characterized in that the hydrophobic, gel-forming or water-insoluble compound comprises a cellulose derivative, polyvinyl alcohol "(PVA) or an edible oil.
14. 14. A granulate according to claim 13, characterized in that the cellulose derivative is hydroxypropylmethylcellulose, carboxymethylcellulose, or hydroxyethylcellulose and / or the edible oil is soybean oil or canola oil.
15. 15. A granulate according to any of claims 9 to 14, characterized in that the enzyme comprises a phytase, endo-xylanase and / or β-glucanase.
16. 16. A process for the preparation of an animal feed, or a premix or precursor for an animal feed, the process is characterized in that it comprises mixing a granulate as defined according to any of claims 9 to 15, with a or more substances or food ingredients for animals.
17. 17. A process according to claim 16, characterized in that the mixture of the food substance (s) and the granulate is sterilized or steam-treated, formed into pellets and optionally dried.
18. 18. A composition, characterized in that it comprises a granulate according to any of claims 9 to 15.
19. 19. A composition according to claim 18, characterized in that it is an edible food composition.
20. 20. A composition according to claim 18 or 19, characterized in that it is an animal feed.
21. 21. A composition according to any of claims 18 to 20, characterized in that it comprises pellets of one or more food substances or food ingredients mixed with a granulate according to any of claims 9 to 15.
22. 22. A composition according to claim 21, characterized in that the proportion of the granulate: food substance (s) or food ingredients is at least 1 g: 1 kg (such as at least 100 ppm).
23. 23. A composition according to any of claims 18 to 22, characterized in that it is an animal feed, or a premix or precursor for an animal feed, preparable by a process according to any of claims 16 or 17.
24. 24. A process for promoting the development of an animal, the process is characterized in that it comprises feeding an animal with a diet comprising either a granulate as defined in any of claims 9 to 15 or a composition as defined according to any of claims 18 to 23.
25. 25. The use of a granulate as defined in any of claims 9 to 15 in, or as a component of, an animal feed or for use in an animal diet.
26. 26. The use of a composition comprising at least 15% (w / w) of an edible carbohydrate polymer as a carrier for an enzyme, to improve the stability of the pellet formation of the enzyme.
27. 27. A process for the preparation of an enzyme-containing granulate, characterized in that a liquid containing enzyme and a solid carrier comprising at least 15% _ (w / w) of starch are mechanically processed to obtain the granules containing enzyme, which They are subsequently dried.