EP1432795A2 - Method for the production of oily suspensions of water-soluble enzymes - Google Patents

Method for the production of oily suspensions of water-soluble enzymes

Info

Publication number
EP1432795A2
EP1432795A2 EP02777104A EP02777104A EP1432795A2 EP 1432795 A2 EP1432795 A2 EP 1432795A2 EP 02777104 A EP02777104 A EP 02777104A EP 02777104 A EP02777104 A EP 02777104A EP 1432795 A2 EP1432795 A2 EP 1432795A2
Authority
EP
European Patent Office
Prior art keywords
water
grinding
soluble
oil
suspension
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP02777104A
Other languages
German (de)
French (fr)
Inventor
Andreas Habich
Frank Runge
Jörg Braun
Preben Soerensen
Wolfgang Heinzl
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
BASF SE
Original Assignee
BASF SE
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by BASF SE filed Critical BASF SE
Priority to EP06120633A priority Critical patent/EP1746155A1/en
Publication of EP1432795A2 publication Critical patent/EP1432795A2/en
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/158—Fatty acids; Fats; Products containing oils or fats
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/174—Vitamins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/22—Compounds of alkali metals
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/24—Compounds of alkaline earth metals, e.g. magnesium
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/20—Inorganic substances, e.g. oligoelements
    • A23K20/28—Silicates, e.g. perlites, zeolites or bentonites
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/20—Shaping or working-up of animal feeding-stuffs by moulding, e.g. making cakes or briquettes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/96—Stabilising an enzyme by forming an adduct or a composition; Forming enzyme conjugates

Definitions

  • the invention relates to oily suspensions of at least one water-soluble enzyme and to a process for the preparation of these suspensions and their use as an additive to animal feed.
  • water-soluble enzymes are mixed into feed in the form of a solid as granules or extrudate before the feed mixture is pelleted.
  • aqueous formulations are commercially available or can be prepared from instant powders by dissolving in water directly Herge ⁇ represents be.
  • Aqueous formulations generally have the noisy part of microbiological stability and low storage stability.
  • JP 09 322 770 describes stable enzyme in oil dispersions and their use for softening meat.
  • This object was achieved according to the invention by a process for the preparation of oily suspensions of water-soluble enzymes, characterized in that in the absence of water-soluble vitamins a) grinding at least one water-soluble enzyme in an oil, preferably in at least one edible oil, down to an average particle size of 0.1 to 100 ⁇ m
  • the water-soluble vitamins are, in particular, ascorbic acid and its salts such as sodium ascorbate and vitamin C derivatives such as sodium, calcium or magnesium ascorbyl-2-monophosphate or calcium ascorbyl-2-polyphosphate, calcium pantothenate , Panthenol, vitamin Bi (thiamine) - as hydrochloride, nitrate or pyrophosphate, vitamin B (riboflavin) and their phosphates, vitamin B 6 and salts, vitamin B ⁇ 2 , biotin, folic acid and folic acid derivatives such as tetrahydrofolic acid, 5-methyltetrahydrofolic acid, 5 -Formyltetrahydrofolic acid, nicotinic acid and nicotinic acid amide.
  • ascorbic acid and its salts such as sodium ascorbate and vitamin C derivatives such as sodium, calcium or magnesium ascorbyl-2-monophosphate or calcium ascorbyl-2-polyphosphate, calcium pantothen
  • Vitamin K 3 (menadione) may also be mentioned as sodium bisulfite as water-soluble vitamin.
  • the water-soluble enzymes are, in particular, oxidoreductases, transferases, lyases, isomerases, ligases and hydrolases.
  • Preferred enzymes are hydrolases. Examples of hydrolase, that is to say enzymes which bring about a hydrolytic cleavage of chemical compounds, esterases, glycosidases, lipases, phospholipases, ether hydrolases, proteases, amidases, aminidases, nitrilases and phosphatases.
  • Glycosidases include both endo- and exo-glucosidases that cleave both ⁇ - and ß-glycosidic bonds. Typical examples of this are: amylases, maltases, cellulases, endo-xylanases, ⁇ -glucanases, mannanases, lysozymes, galactosidases, ⁇ -glucuronidases, glucose oxidases, saccharases, lactases and the like.
  • Non-starch polysaccharide-cleaving enzymes such as, for example, amylase, glucanase and xylanase, and phosphatases, such as, in particular, phytase are particularly preferred.
  • the water-soluble enzymes can also be used as a combination of the enzymes described.
  • the embodiments described below therefore always relate to the combination of water-soluble enzymes.
  • the above-mentioned water-soluble enzymes can be in any form in solid form before grinding. These solids can be produced in a manner known per se, for example by drying or precipitation processes from culture broths. These culture broths are preferably filtered or centrifuged before drying or precipitation. Drying processes are, for example, spray drying, fluidized bed drying or contact drying processes, in particular freeze drying processes.
  • the precipitation processes are known per se, for example precipitation by addition of Na 2 SO 4 .
  • Precipitation methods are described, for example, in "Enzyme_ der Tierernahrung, AWT, 1997, Roonstrasse 53175 Bonn”.
  • the water-soluble enzymes can be present, for example, as powders, granules or lyophyllisates.
  • the water-soluble enzymes are preferably used as solids which have a water content of less than 10% by weight.
  • Edible oils are generally all physiologically acceptable oils - both vegetable and animal - in question, in particular those oils which are liquid at 20 ° C or in the suspension at 20 ° C alone or together with other oils form the liquid phase.
  • Sunflower oil, palm oil, sesame oil, corn oil, cottonseed oil, soybean oil or peanut oil, esters of medium-chain triglycerides and also fish oils such as, for example, mackerel, sprat or salmon oil are preferred in this connection.
  • Fish oils, corn oil, sunflower oil and peanut oil are particularly preferred for animal nutrition.
  • the esters of medium-chain triglycerides are also advantageous for the food / pharmaceutical sector.
  • Vitamin E, vitamin E derivatives or mixtures thereof are also to be understood as edible oil in the sense of the invention.
  • the term vitamin E in this context stands for natural or synthetic ot-, ß-, ⁇ - or ⁇ -tocopherol, preferably for natural or synthetic tocopherol and for tocotrienol.
  • Vitamin E derivatives are, for example, tocopheryl-C-co-alkanoic acid esters such as tocopheryl acetate or tocopheryl palmitate. Vitamin E and / or their derivatives can be used alone or together with the other edible oils as a dispersing medium.
  • the grinding can be carried out in a manner known per se e.g. done with a ball mill. Depending on the type of mill used, grinding is carried out until the particles have an average particle size D [4.3] of 0.1 to 100 ⁇ m, preferably 0.2 to 80 ⁇ m, particularly preferably 0.5 to 50 ⁇ m, very particularly preferably 0.8 to 40 ⁇ m. , measured for example via Fraunhofer diffraction.
  • D [4,3] denotes the volume-weighted mean diameter (see manual for Malvern Mastersizer S, Malvern Instruments Ltd., UK).
  • each individual component to be ground can also be ground in high concentration in the oil to be used.
  • the final preparation then results from a mixture of the individual suspensions.
  • the preparation according to the invention can be diluted with fats or oils to the respective use concentration before use.
  • a special embodiment of the process according to the invention is characterized in that the grinding in step a) and the grinding and / or suspension in step b) are carried out in the absence of an emulsifier.
  • a further advantageous embodiment of the method according to the invention is characterized in that the grinding in step a TM) and the grinding and / or suspending in step b effected) in the absence of a protective colloid.
  • the per se hydrophilic enzymes unexpectedly for the person skilled in the art - could be finely ground without wetting problems and agglomerate formation in the above-mentioned hydrophobic dispersing media. 5
  • oily suspensions according to the invention can also be prepared by dry grinding the water-soluble enzymes and then suspending the ground particles in at least one edible oil. 10
  • dry grinding means grinding without the use of a continuous phase.
  • Desiccants selected from the group consisting of alkali and alkaline earth metal sulfates such as sodium, calcium and magnesium sulfate, alkali and alkaline earth metal chlorides such as sodium, calcium and magnesium chloride and silica gel are preferred. Desiccant is very particularly preferred Call CaCl 2
  • the amount of desiccant used is generally between 0.1 and 20% by weight, preferably between 0.5 and 15% by weight, particularly preferably between 1.0 and 10% by weight, based on the total amount of oily suspension.
  • the one or more desiccant used can thereby also separately - as in the method step a) - milled in an edible oil 'advertising the 5 and then the oily suspension of the ground water-soluble enzymes are added. It is also possible to mix the desiccants even unground with the oily suspension of the ground water-soluble enzymes from process step a). In the case of dry grinding, the water-soluble enzymes and the drying agent (s) can also be ground separately " and then added to the oily suspension.
  • the addition of desiccants brings about a reduction in the viscosity of the oily suspension in comparison with an oily suspension free of desiccants. Because of the finely divided distribution of the dispersed water-soluble enzymes, the oily suspensions produced by the process according to the invention are distinguished by a high bioavailability of the active ingredients contained in the suspension. 5
  • fat-soluble vitamins such as the K vitamins, vitamin A and derivatives such as vitamin A acetate, vitamin A propionate or vitamin A palmitate, vitamin D 2 10 and Vitamin D and the aforementioned E vitamins are added to the oily suspension and dissolved.
  • the grinding in step a) and the suspension in step b) are preferably carried out in the presence of fat-soluble vitamins.
  • the invention also relates to oily suspensions of at least one water-soluble enzyme, obtainable by the process according to the invention described above.
  • the oily suspensions according to the invention contain 1 to 20 70% by weight, preferably 2 to 60% by weight, particularly preferably 10 to 55% by weight, very particularly preferably 15 to 50% by weight of at least one of the water-soluble enzymes mentioned at the outset in finely ground form.
  • oily suspensions can additionally 0.5 to
  • oily preparations can additionally contain at least one further carotenoid.
  • Carotenoids are e.g. to understand the following compounds: 35 ß-carotene, lycopene, lutein, astaxanthin, zeaxanthin, cryptoxanthine, citranaxanthin, canthaxanthin, bixin, ß-apo-4-carotinal, ß-apo-8-carotinal, ß-apo-8- carotenic acid esters, individually or as a mixture.
  • Carotenoids which are preferably used are ⁇ -carotene, lycopene, lutein, astaxanthin, zeaxanthin, citranaxanthin and canthaxanthin. ⁇
  • the carotenoids can be used in crystalline form or as a formulation - for example as dry powder, according to EP-A-0 065 193. 45
  • the carotenoids are usually ground in crystalline form together with the water-soluble enzymes in the oil.
  • the content of carotenoids in the formulations is generally between 0.1 and 40% by weight, preferably between 0.3 and 20% by weight, particularly preferably between 0.5 and 10% by weight, very particularly preferably between 1 and 5 wt .-%, based on the total amount of the formulation.
  • oily preparations according to the invention can contain up to 10% by weight of additional components such as minerals, amino acids, proteins or fat-soluble enzymes.
  • fat-soluble vitamins and carotenoids can be added before, during or after the grinding of the suspension according to the invention.
  • Grind additives In order to obtain a homogeneous suspension of all non-oil-soluble components that is as fine as possible, it is advantageous to use the above-mentioned Grind additives together with the water-soluble enzymes.
  • minerals for example iron sulfate, zinc sulfate, manganese sulfate, copper sulfate, calcium sulfate, sodium sulfate, copper oxide, magnesium oxide, calcium fluoride, potassium chloride, potassium iodide, sodium chloride, calcium iodate, calcium, magnesium, potassium, sodium or iron phosphate, cobalt carbonate, Sodium selenate or silica and their salts are incorporated into the suspension and also ground.
  • the amount of minerals used, for example in the animal nutrition sector, is based on the needs of the animals to be fed.
  • All known physiologically harmless ⁇ -amino acid residues are generally suitable as amino acid residues.
  • the residues of the following amino acids are preferred: alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, hippuric acid, serine and taurine. Lysine, methionine and cysteine are particularly preferred.
  • compositions with a vitamin or coenzyme character e.g. B. Choline chloride, carnitine, ⁇ -butyrobetaine, lipoic acid, creatine, ubiquinones, S-methyl methionine, S-adenosyl methionine.
  • Polyunsaturated fatty acids e.g. B. Linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid.
  • oily suspensions also contain adjuvants such as e.g. Protective colloids, antioxidants, thickeners, chelating agents, such as e.g. Contain alkali or alkaline earth salts of citric acid, phytic acid or phosphoric acid and / or contain emulsifiers.
  • adjuvants such as e.g. Protective colloids, antioxidants, thickeners, chelating agents, such as e.g. Contain alkali or alkaline earth salts of citric acid, phytic acid or phosphoric acid and / or contain emulsifiers.
  • Protective colloids that can be used are, for example, gelatin, fish gelatin, starch, dextrin, plant proteins, pectin, gum arabic, casein, caseinate or mixtures thereof.
  • polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, carboxymethyl cellulose, hydroxypropyl cellulose and alginates can also be used.
  • stabilizers such as ⁇ -tocopherol, t-butyl-hydroxy-toluene, t-butylhydroxyanisole or ethoxyquin.
  • emulsifiers or solubilizers which can be used are polyglycerol fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters or lecithin.
  • the suspensions are u. a. as an additive in animal feed preparations or in compound feed and for the production of nutritional supplements in the animal sector.
  • the suspensions can preferably be used as a feed additive in animal nutrition, preferably for application or "spraying" onto feed pellets, particularly preferably for admixing with feed mixtures before pelleting. It is used as a feed additive in particular by spraying the suspensions according to the invention directly, optionally after dilution with oils, for example on animal feed pellets as a so-called "post-pelleting application”.
  • a preferred embodiment of the spraying process consists in loading the feed pellets with the oily suspension under reduced pressure.
  • Examples include: in GB-A-2 232 573 and in EP-A-0 556 883.
  • the invention is preferably directed to animal feed, in particular feed pellets which are loaded with the suspensions or are mixed in desired combinations before the feed mixtures are pelletized.
  • the oily suspensions which can be produced by the process according to the invention have the advantage of increased storage stability and reduced susceptibility to microbiological contamination.
  • oily suspensions can be added directly to putter mixes prior to pelleting without the complex procedural step of the coating.
  • Two kilograms of a mixture of 25% by weight of dried phytase (from Asperg-illus, spray-dried) and 75% by weight of corn oil are stirred with a blade stirrer until a homogeneous suspension is obtained.
  • the mixture is then poured into a stirrable receiver, from which the suspension is conveyed by means of a peristaltic pump through a continuously operated ball mill (Dyno Mil-TKDL Special).
  • the grinding bowl of the ball mill is filled with 400 g glass balls (diameter 800 to 1200 ⁇ m).
  • the fine-particle suspension emerging from the mill is collected and measured using a particle size measuring device (Malvern Mastersizer). The grinding process is repeated until the suspended particles have an average particle size of less than 20 ⁇ m.

Abstract

The invention relates to oily suspensions of at least one water-soluble enzyme and to a method for the production of said suspensions and the use thereof as additives for animal feed.

Description

Verfahren zur Herstellung öliger Suspensionen wasserlöslicher En¬ zymeA process for the preparation of water-soluble oily suspensions En ¬ zyme
Beschreibungdescription
Die Erfindung betrifft ölige Suspensionen mindestens eines wasserlöslichen Enzyms sowie ein Verfahren zur Herstellung dieser Suspensionen und deren Verwendung als Zusatz zu Tierfuttermit- teln.The invention relates to oily suspensions of at least one water-soluble enzyme and to a process for the preparation of these suspensions and their use as an additive to animal feed.
Das Zumischen von wasserlöslichen Enzymen zu Futtermitteln erfolgt im Stand der Technik als Feststoff als Granulat oder Extru- dat vor dem Pelletieren der Futtermittelmischung.In the prior art, water-soluble enzymes are mixed into feed in the form of a solid as granules or extrudate before the feed mixture is pelleted.
Dies hat den Nachteil, daß zum Erreichen ausreichender Pelletier¬ stabilität bei höheren Pelletiertemperaturen das Aufbringen von Umhüllungen auf d s Granulat oder Extrudat in einem zusätzlichen Verfahrensschritt erforderlich ist.This has the disadvantage that to achieve sufficient stability Pelletier ¬ at higher pelleting temperatures the application of wrappers on ds granules or extrudate required in an additional process step.
Weiterhin können flüßige, wäßrige Formulierungen von wasserlöslichen Enzymen nach dem Pelletieren durch Aufsprühen auf die Fut¬ termittelpellets θppliziert werden.Can continue to liquid aqueous formulations of water-soluble enzymes after pelleting by spraying onto the Fut ¬ termittelpellets be θppliziert.
Diese wäßrigen Formulierungen sind kommerziell Erhältlich oder können aus Instantpulvern durch Lösen in Wasser direkt herge¬ stellt werden.These aqueous formulations are commercially available or can be prepared from instant powders by dissolving in water directly Herge ¬ represents be.
Wäßrige Formulierungen weisen in der Regel den Krachteil mikrobio- logischer Stabilität und eine geringe Lagerstabilität auf.Aqueous formulations generally have the noisy part of microbiological stability and low storage stability.
JP 09 322 770 beschreibt stabile Enzym in Öl-Dis;persionen und deren Verwendung zum Weichen von Fleisch.JP 09 322 770 describes stable enzyme in oil dispersions and their use for softening meat.
Es war daher die Aufgabe der vorliegenden Erfindung, stabile flüssige Formulierungen wasserlöslicher Enzyme bereitzustellen, die die oben genannten Nachteile des Standes der Technik nicht aufweisen.It was therefore the object of the present invention to provide stable liquid formulations of water-soluble enzymes which do not have the above-mentioned disadvantages of the prior art.
Diese Aufgabe wurde erfindungsgemäß gelöst durch ein Verfahren zur Herstellung öliger Suspensionen wasserlöslicher Enzyme, dadurch gekennzeichnet, daß man in Abwesenheit wasserlöslicher Vitamine a) mindestens ein wasserlösliches Enzym in einem Öl, bevorzugt in mindestens einem eßbaren Öl bis auf eine mittlere Partikelgröße von 0,1 bis 100 Um mahlt oderThis object was achieved according to the invention by a process for the preparation of oily suspensions of water-soluble enzymes, characterized in that in the absence of water-soluble vitamins a) grinding at least one water-soluble enzyme in an oil, preferably in at least one edible oil, down to an average particle size of 0.1 to 100 μm
b) mindestens ein wasserlösliches Enzym ohne Verwendung einer kontinuierlichen Phase bis auf eine mittlere Partikelgröße von 0,1 bis 100 um mahlt und die gemahlenen Partikel anschließend in einem Öl, bevorzugt in mindestens einem eßbaren Öl suspendiert .b) grinding at least one water-soluble enzyme without using a continuous phase to an average particle size of 0.1 to 100 μm and then suspending the ground particles in an oil, preferably in at least one edible oil.
Bei den wasserlöslichen Vitaminen handelt es sich insbesondere um Ascorbinsäure und deren Salze wie Natriumascorbat sowie Vitamin C-Derivate wie Natrium-, Calcium- oder Magnesium-ascorbyl-2-mono- phosphat oder Calcium-ascorbyl-2-polyphosphat, Calcium-panto- thenat, Panthenol, Vitamin Bi (Thiamin) - als Hydrochlorid, Nitrat oder Pyrophosphat , Vitamin B (Riboflavin) und deren Phosphate, Vitamin B6 und Salze, Vitamin Bι2, Biotin, Folsäure und Folsäure- derivate wie Tetrahydrofolsäure, 5-Methyltetrahydrofolsäure, 5-Formyltetrahydrofolsäure, Nicotinsäure und Nicotinsäureamid.The water-soluble vitamins are, in particular, ascorbic acid and its salts such as sodium ascorbate and vitamin C derivatives such as sodium, calcium or magnesium ascorbyl-2-monophosphate or calcium ascorbyl-2-polyphosphate, calcium pantothenate , Panthenol, vitamin Bi (thiamine) - as hydrochloride, nitrate or pyrophosphate, vitamin B (riboflavin) and their phosphates, vitamin B 6 and salts, vitamin Bι 2 , biotin, folic acid and folic acid derivatives such as tetrahydrofolic acid, 5-methyltetrahydrofolic acid, 5 -Formyltetrahydrofolic acid, nicotinic acid and nicotinic acid amide.
Als wasserlösliches Vitamin sei in diesem Zusammenhang auch Vitamin K3 (Menadion) als Natriumbisulfit genannt.Vitamin K 3 (menadione) may also be mentioned as sodium bisulfite as water-soluble vitamin.
Bei den wasserlöslichen Enzymen handelt es sich insbesondere Oxi- doreduktasen, Transferasen, Lyasen, Isomerasen, Ligasen und Hy- drolasen. Bevorzugte Enzyme sind Hydrolasen. Beispiele für Hydro- lase, daß heißt Enzyme welche eine hydrolytische Spaltung von chemischen Verbindungen bewirken sind, Esterasen, Glycosidasen, Lipasen, Phospholipasen, Etherhydrolasen, Proteasen, Amidasen, Aminidasen, Nitrilasen und Phosphatasen.The water-soluble enzymes are, in particular, oxidoreductases, transferases, lyases, isomerases, ligases and hydrolases. Preferred enzymes are hydrolases. Examples of hydrolase, that is to say enzymes which bring about a hydrolytic cleavage of chemical compounds, esterases, glycosidases, lipases, phospholipases, ether hydrolases, proteases, amidases, aminidases, nitrilases and phosphatases.
Glycosidasen umfassen sowohl endo- als auch exo-Glucosidasen, die sowohl α- als auch ß-glycosidische Bindungen spalten. Typische Beispiele hierfür sind: Amylasen, Maltasen, Cellulasen, Endo-Xy- lanasen, ß-Glucanasen, Mannanasen, Lysozyme, Galactosidasen, ß-Glucuronidasen, Glucoseoxidasen, Saccharasen, Lactasen und der- geleichen.Glycosidases include both endo- and exo-glucosidases that cleave both α- and ß-glycosidic bonds. Typical examples of this are: amylases, maltases, cellulases, endo-xylanases, β-glucanases, mannanases, lysozymes, galactosidases, β-glucuronidases, glucose oxidases, saccharases, lactases and the like.
Bevorzugt sind insbesondere nichtstärkepolysaccharidspaltende En- zyme, wie zum Beispiel Amylase, Glucanase und Xylanase sowie Phosphatasen, wie insbesondere Phytase.Non-starch polysaccharide-cleaving enzymes, such as, for example, amylase, glucanase and xylanase, and phosphatases, such as, in particular, phytase are particularly preferred.
Im erfindungsgemäßen Verfahren können die wasserlöslichen Enzyme auch als Kombination der beschriebenen Enzyme eingesetzt werden. Die nachstehend beschriebenen Ausführungsformen beziehen sich daher immer auch auf die Kombination von wasserlöslichen Enzymen.In the process according to the invention, the water-soluble enzymes can also be used as a combination of the enzymes described. The embodiments described below therefore always relate to the combination of water-soluble enzymes.
Die oben genannten wasserlöslichen Enzyme können vor der Mahlung in beliebiger Weise in fester Form vorliegen. Herstellbar sind diese Feststoffe in an sich bekannter Weise beispielsweise durch Trocknungs- oder Fällungsverfahren aus Kulturbrühen. Vorzugsweise werden diese Kulturbrühen vor der Trocknung oder Fällung filtriert oder zentrifugiert . Trocknungsverfahren sind beispiels- weise Sprühtrocknungs-, Wirbelschichttrocknungs- oder Kontakttrocknungsverfahren, insbesondere Gefriertrocknungsverfahren.The above-mentioned water-soluble enzymes can be in any form in solid form before grinding. These solids can be produced in a manner known per se, for example by drying or precipitation processes from culture broths. These culture broths are preferably filtered or centrifuged before drying or precipitation. Drying processes are, for example, spray drying, fluidized bed drying or contact drying processes, in particular freeze drying processes.
Die Fällungsverfahren sind an sich bekannt, wie beispielsweise Fällung durch Na2Sθ4-Zusatz . Fällungsmethoden sind beispielsweise in "Enzyme_ der Tierernährung, AWT, 1997, Roonstraße 53175 Bonn" beschrieben.The precipitation processes are known per se, for example precipitation by addition of Na 2 SO 4 . Precipitation methods are described, for example, in "Enzyme_ der Tierernahrung, AWT, 1997, Roonstrasse 53175 Bonn".
Dementsprechend können die wasserlöslichen Enzyme beispielsweise als Pulver, Granulate oder Lyophyllisate vorliegen.Accordingly, the water-soluble enzymes can be present, for example, as powders, granules or lyophyllisates.
Bevorzugt werden die wasserlöslichen Enzyme vor der Mahlung als Feststoffe verwendet, die einen Wassergehalt kleiner 10 Gew.-% aufweisen.Before the grinding, the water-soluble enzymes are preferably used as solids which have a water content of less than 10% by weight.
Als eßbare Öle kommen in der Regel alle physiologisch unbedenklichen 'Öle - sowohl pflanzlichen als auch tierischen Ursprungs - in Frage, insbesondere solche Öle, die bei 20°C flüssig sind bzw. die in der Suspension bei 20°C allein oder zusammen mit anderen Ölen die flüssige Phase bilden. Bevorzugt zu nennen sind in diesem Zusammenhang Sonnenblumenöl, Palmöl, Sesamöl, Maiskei- möl, Baumwollsaatöl, So abohnenöl oder Erdnußöl, Ester mittel- kettiger Triglyceride sowie außerdem Fischöle wie beispielsweise Makrelen-, Sprotten- oder Lachsöl. Für die Tierernährung besonders bevorzugt sind Fischöle, Maiskeimöl, Sonnenblumenöl und Erdnußöl. Für den Food-/Pharmabereich zusätzlich von Vorteil sind die Ester mittelkettiger Triglyceride.Edible oils are generally all physiologically acceptable oils - both vegetable and animal - in question, in particular those oils which are liquid at 20 ° C or in the suspension at 20 ° C alone or together with other oils form the liquid phase. Sunflower oil, palm oil, sesame oil, corn oil, cottonseed oil, soybean oil or peanut oil, esters of medium-chain triglycerides and also fish oils such as, for example, mackerel, sprat or salmon oil are preferred in this connection. Fish oils, corn oil, sunflower oil and peanut oil are particularly preferred for animal nutrition. The esters of medium-chain triglycerides are also advantageous for the food / pharmaceutical sector.
Als eßbares Öl im Sinne der Erfindung sind auch Vitamin E, Vitamin E-Derivate oder Mischungen davon zu verstehen. Die Be- Zeichnung Vitamin E steht in diesem Zusammenhang für natürliches oder synthetisches ot-, ß- , γ- oder δ-Tocopherol, bevorzugt für natürliches oder synthetisches -Tocopherol sowie für Toco- trienol. Vitamin E-Derivate sind z.B. Tocopheryl-C-Co-Alkansäu- reester wie Tocopherylacetat oder Tocopherylpalmitat . Vitamin E und/oder deren Derivate können dabei allein oder zusammen mit den anderen eßbaren Ölen als Dispergiermedium verwendet werden.Vitamin E, vitamin E derivatives or mixtures thereof are also to be understood as edible oil in the sense of the invention. The term vitamin E in this context stands for natural or synthetic ot-, ß-, γ- or δ-tocopherol, preferably for natural or synthetic tocopherol and for tocotrienol. Vitamin E derivatives are, for example, tocopheryl-C-co-alkanoic acid esters such as tocopheryl acetate or tocopheryl palmitate. Vitamin E and / or their derivatives can be used alone or together with the other edible oils as a dispersing medium.
Die Mahlung kann in an sich bekannter Weise z.B. mit einer Kugelmühle erfolgen. Dabei wird je nach verwendetem Mühlentyp so lange gemahlen, bis die Teilchen eine mittlere Partikelgröße D[4,3] von 0,1 bis 100 μm, bevorzugt 0,2 bis 80 μm, besonders bevorzugt 0,5 bis 50 μ , ganz besonders bevorzugt 0,8 bis 40 μm. , beispielsweise über Fraunhofer Beugung gemessen, aufweisen. Der Begriff D[4,3] bezeichnet den volumengewichteten mittleren Durchmesser (siehe Handbuch zu Malvern Mastersizer S, Malvern Instruments Ltd. , UK) .The grinding can be carried out in a manner known per se e.g. done with a ball mill. Depending on the type of mill used, grinding is carried out until the particles have an average particle size D [4.3] of 0.1 to 100 μm, preferably 0.2 to 80 μm, particularly preferably 0.5 to 50 μm, very particularly preferably 0.8 to 40 μm. , measured for example via Fraunhofer diffraction. The term D [4,3] denotes the volume-weighted mean diameter (see manual for Malvern Mastersizer S, Malvern Instruments Ltd., UK).
Es kann vorteilhaft sein, bei großen Festkörpern eine Vormahlung der Enzymfeststoffe durchzuführen.It may be advantageous to pre-grind the enzyme solids for large solids.
Nähere Einzelheiten zur Mahlung und den dafür eingesetzten Apparaturen finden sich u.a: in Ullmannxs Encyclopedia of Industrial Chemistry, Sixth Edition, 1999, Electronic Release, Size Reduction, Kapitel 3.6.: Wet Grinding.Further details on grinding and the equipment used for this can be found, inter alia: in Ullmann x s Encyclopedia of Industrial Chemistry, Sixth Edition, 1999, Electronic Release, Size Reduction, Chapter 3.6 .: Wet Grinding.
Bei dem erfindungsgemäßen Mahlverfahren ist es möglich, alle in der Suspension verwendeten Komponenten als Gesamtmischung zu mah- len. Es kann aber auch jede einzelne zu mahlende Komponente in hoher Konzentration in dem zu verwendenden Öl gemahlen werden. Die Endzubereitung ergibt sich dann durch eine Abmischung der jeweiligen Einzelsuspensionen.In the grinding process according to the invention, it is possible to grind all of the components used in the suspension as an overall mixture. However, each individual component to be ground can also be ground in high concentration in the oil to be used. The final preparation then results from a mixture of the individual suspensions.
Die erfindungsgemäße Zubereitung kann vor der Anwendung mit Fetten oder Ölen auf die jeweilige Gebrauchskonzentration verdünnt werden.The preparation according to the invention can be diluted with fats or oils to the respective use concentration before use.
Eine besondere Ausführungsform des erfindungsgemäßen Verfahrens ist dadurch gekennzeichnet, daß die Mahlung im Schritt a) sowie die Mahlung und/oder Suspendierung im Schritt b) in Abwesenheit eines Emulgators erfolgen.A special embodiment of the process according to the invention is characterized in that the grinding in step a) and the grinding and / or suspension in step b) are carried out in the absence of an emulsifier.
Eine weitere vorteilhafte Ausführungsform des erfindungsgemäßen Verfahrens ist dadurch gekennzeichnet, daß die Mahlung im™Schritt a) sowie die Mahlung und/oder Suspendierung im Schritt b) in Abwesenheit eines Schutzkolloids erfolgen. Trotz Fehlen der o.g. Dispergier- bzw. Formulierhilfsmittel konnten die an sich hydrophilen Enzyme - für den Fachmann unerwartet - ohne Benetzungsprobleme und Agglomeratbildung in den o.g. hydrophoben Dispergiermedien feinstgemahlen werden. 5A further advantageous embodiment of the method according to the invention is characterized in that the grinding in step a ™) and the grinding and / or suspending in step b effected) in the absence of a protective colloid. Despite the absence of the above-mentioned dispersing or formulation auxiliaries, the per se hydrophilic enzymes - unexpectedly for the person skilled in the art - could be finely ground without wetting problems and agglomerate formation in the above-mentioned hydrophobic dispersing media. 5
Neben der oben beschriebenen Naßmahlung, lassen sich die erfindungsgemäßen öligen Suspensionen auch durch Trockenmahlung der wasserlöslichen Enzyme und anschließendes Suspendieren der gemahlenen Partikel in mindestens einem eßbaren Öl herstellen. 10 Als Trockenmahlung versteht man in diesem Zusammenhang eine Mahlung ohne Verwendung einer kontinuierlichen Phase.In addition to the wet grinding described above, the oily suspensions according to the invention can also be prepared by dry grinding the water-soluble enzymes and then suspending the ground particles in at least one edible oil. 10 In this context, dry grinding means grinding without the use of a continuous phase.
Nähere Einzelheiten zur Trockenmahlung finden sich u.a. in Ullmann's Encyclopedia of Industrial Chemistry, Sixth Edition, 15 1999, Electronic Release, Size Reduction, Kapitel 3.4.Further details on dry grinding can be found, among others, in Ullmann's Encyclopedia of Industrial Chemistry, Sixth Edition, 15 1999, Electronic Release, Size Reduction, Chapter 3.4.
Als besonderen Vorteil hinsichtlich der Stabilität der erfindungsgemäßen öligen Dispersionen hat sich herausgestellt, wenn man die Mahlung imVerfahrensschritt a) sowie die MahlungA particular advantage with regard to the stability of the oily dispersions according to the invention has been found when the grinding in process step a) and the grinding
20 und/oder Suspendierung im Schritt b) in Gegenwart von Trockenmitteln durchführt. Bevorzugt sind dabei Trockenmittel, ausgewählt aus der Gruppe, bestehend aus Alkali- und Erdalkalisulfaten wie Natrium-, Calcium- und Magnesiumsulfat, Alkali- und Erdalkalichloriden wie Natrium-, Calcium- und Magnesiumchlorid und Kiesel- 5 gel. Als- ganz besonders bevorzugtes Trockenmittel ist CaCl2 zu nenne20 and / or suspension in step b) in the presence of drying agents. Desiccants selected from the group consisting of alkali and alkaline earth metal sulfates such as sodium, calcium and magnesium sulfate, alkali and alkaline earth metal chlorides such as sodium, calcium and magnesium chloride and silica gel are preferred. Desiccant is very particularly preferred Call CaCl 2
Die Menge an eingesetztem Trockenmittel liegt im allgemeinen zwischen 0,1 und 20 Gew.-%, bevorzugt zwischen 0,5 und 15 Gew.-%, 0 besonders bevorzugt zwischen 1,0 und 10 Gew.-%, bezogen auf die Gesamtmenge der öligen Suspension.The amount of desiccant used is generally between 0.1 and 20% by weight, preferably between 0.5 and 15% by weight, particularly preferably between 1.0 and 10% by weight, based on the total amount of oily suspension.
Das oder die verwendeten Trockenmittel können dabei auch separat - wie im Verfahrensschritt a) - in einem eßbaren Öl gemahlen' wer- 5 den und anschließend der öligen Suspension der gemahlenen wasserlöslichen Enzyme zugesetzt werden. Ferner ist es möglich, die Trockenmittel auch ungemahlen mit der öligen Suspension der gemahlenen wasserlöslichen Enzyme aus Verfahrensschritt a) zu mischen. Im Falle der Trockenmahlung können die wasserlöslichen En- 0 zyme und das oder die Trockenmittel auch separat gemahlen"und dann zur öligen Suspension gegeben werden.The one or more desiccant used can thereby also separately - as in the method step a) - milled in an edible oil 'advertising the 5 and then the oily suspension of the ground water-soluble enzymes are added. It is also possible to mix the desiccants even unground with the oily suspension of the ground water-soluble enzymes from process step a). In the case of dry grinding, the water-soluble enzymes and the drying agent (s) can also be ground separately " and then added to the oily suspension.
Es wird überraschenderweise gefunden, daß die Zugabe von Trockenmitteln eine Verminderung der Viskosität der öligen Suspension im 5 Vergleich zu trockenmittelfreien öligen- Suspension bewirkt. Aufgrund der feinteiligen Verteilung der dispergierten wasserlöslichen Enzyme zeichnen sich die nach dem erfindungsgemäßen Verfahren hergestellten öligen Suspensionen durch eine hohe Bioverfügbarkeit der in der Suspension enthaltenen Wirkstoffe aus . 5It is surprisingly found that the addition of desiccants brings about a reduction in the viscosity of the oily suspension in comparison with an oily suspension free of desiccants. Because of the finely divided distribution of the dispersed water-soluble enzymes, the oily suspensions produced by the process according to the invention are distinguished by a high bioavailability of the active ingredients contained in the suspension. 5
Neben den eingangs genannten wasserlöslichen Enzymen können vor, während oder nach der Mahlung zusätzliche fettlösliche Vitamine, wie z.B. die K-Vitamine, Vitamin A und Derivate wie Vitamin A- Acetat, Vitamin A-Propionat oder Vitamin A-Palmitat, Vitamin D2 10 und Vitamin D sowie die bereits genannten E-Vitamine in die ölige Suspension eingetragen und gelöst werden. Bevorzugt erfolgt die Mahlung im Schritt a) sowie die Suspendierung im Schritt b) in Gegenwart fettlöslicher Vitamine.In addition to the water-soluble enzymes mentioned at the beginning, additional fat-soluble vitamins such as the K vitamins, vitamin A and derivatives such as vitamin A acetate, vitamin A propionate or vitamin A palmitate, vitamin D 2 10 and Vitamin D and the aforementioned E vitamins are added to the oily suspension and dissolved. The grinding in step a) and the suspension in step b) are preferably carried out in the presence of fat-soluble vitamins.
15 Gegenstand_ der Erfindung sind auch ölige Suspensionen mindestens eines wasserlöslichen Enzyms, erhältlich nach dem erfindungsgemäßen, vorstehend beschriebenen Verfahren.The invention also relates to oily suspensions of at least one water-soluble enzyme, obtainable by the process according to the invention described above.
Die erfindungsgemäßen öligen Suspensionen enthalten dabei 1 bis 20 70 Gew.-%, bevorzugt 2 bis 60 Gew.-%, besonders bevorzugt 10 bis 55 Gew.-% ganz besonders bevorzugt 15 bis 50 Gew.-% mindestens eines der eingangs genannten wasserlöslichen Enzyme in feinstver- mahlener Form.The oily suspensions according to the invention contain 1 to 20 70% by weight, preferably 2 to 60% by weight, particularly preferably 10 to 55% by weight, very particularly preferably 15 to 50% by weight of at least one of the water-soluble enzymes mentioned at the outset in finely ground form.
25 Außerdem können die öligen Suspensionen zusätzlich 0,5 bis25 In addition, the oily suspensions can additionally 0.5 to
60 Gew -%, bevorzugt 5 bis 50 Gew.-%, besonders bevorzugt 10 bis 45 Gew.-%, ganz besonders bevorzugt 15 bis 40 Gew.-% mindestens eines der eingangs genannten fettlöslichen Vitamine in gelöster Form enthalten.60% by weight, preferably 5 to 50% by weight, particularly preferably 10 to 45% by weight, very particularly preferably 15 to 40% by weight, of at least one of the fat-soluble vitamins mentioned at the outset in dissolved form.
3030
Darüberhinaus können die öligen Zubereitungen zusätzlich mindestens ein weiteres Carotinoid enthalten.In addition, the oily preparations can additionally contain at least one further carotenoid.
Unter Carotinoide sind z.B. folgende Verbindungen zu verstehen: 35 ß-Carotin, Lycopin, Lutein, Astaxanthin, Zeaxanthin, Crypto- xanthin, Citranaxanthin, Canthaxanthin, Bixin, ß-Apo-4-carotinal, ß-Apo-8-carotinal, ß-Apo-8-carotinsäureester, einzeln oder als Mischung. Bevorzugt verwendete Carotinoide sind ß-Carotin, Lycopin, Lutein, Astaxanthin, Zeaxanthin, Citranaxanthin und 40 Canthaxanthin. ~~Carotenoids are e.g. to understand the following compounds: 35 ß-carotene, lycopene, lutein, astaxanthin, zeaxanthin, cryptoxanthine, citranaxanthin, canthaxanthin, bixin, ß-apo-4-carotinal, ß-apo-8-carotinal, ß-apo-8- carotenic acid esters, individually or as a mixture. Carotenoids which are preferably used are β-carotene, lycopene, lutein, astaxanthin, zeaxanthin, citranaxanthin and canthaxanthin. ~~
Die Carotinoide können dabei in kristalliner Form oder als Formulierung - beispielsweise als Trockenpulver, gemäß EP-A-0 065 193 eingesetzt werden. 45 Vorteilhafterweise werden die Carotinoide in der Regel in kristalliner Form zusammen mit den wasserlöslichen Enzymen in dem Öl gemahlen. Im Falle von Astaxanthin und Canthaxanthin werden bevorzugt Astaxanthin- bzw. Canthaxanthin-haltige Trockenpulver, beispielsweise Lucantin Pink bzw. Lucantin Rot (ein 10%iges Astaxanthin- bzw. Canthaxanthin-Trockenpulver, Fa. BASF Aktiengesellschaft, Ludwigshafen, Deutschland) zusammen mit den wasserlöslichen Enzymen eingesetzt.The carotenoids can be used in crystalline form or as a formulation - for example as dry powder, according to EP-A-0 065 193. 45 Advantageously, the carotenoids are usually ground in crystalline form together with the water-soluble enzymes in the oil. In the case of astaxanthin and canthaxanthin, preference is given to dry powders containing astaxanthin or canthaxanthin, for example Lucantin Pink or Lucantin Red (a 10% strength astaxanthin or canthaxanthin dry powder, from BASF Aktiengesellschaft, Ludwigshafen, Germany) together with the water-soluble Enzymes used.
Der Gehalt an Carotinoide liegt in den Formulierungen im allgemeinen zwischen 0,1 und 40 Gew.-%, bevorzugt zwischen 0,3 und 20 Gew.-%, besonders bevorzugt zwischen 0,5 und 10 Gew.-%, ganz besonders bevorzugt zwischen 1 und 5 Gew.-%, bezogen auf die Gesamtmenge der Formulierung.The content of carotenoids in the formulations is generally between 0.1 and 40% by weight, preferably between 0.3 and 20% by weight, particularly preferably between 0.5 and 10% by weight, very particularly preferably between 1 and 5 wt .-%, based on the total amount of the formulation.
Je nach Anwendungszweck können die erfindungsgemäßen öligen Zubereitungen bis zu 10 Gew.-% weiterer Zusatzkomponenten wie beispielsweise Mineralstoffe, Aminosäuren, Proteine oder fettlösliche Enzyme.Depending on the intended use, the oily preparations according to the invention can contain up to 10% by weight of additional components such as minerals, amino acids, proteins or fat-soluble enzymes.
Diese Zusatzstoffe, ebenso wie die o.g. fettlöslichen Vitamine und Carotinoide, können vor, während oder nach der Mahlung der erfindungsgemäßen Suspension zugesetzt werden. Um eine möglichst feinteilige homogene Suspension aller nicht-öllöslicher Bestand- teile zu- erhalten, ist es von Vorteil, die o.g. Zusatzstoffe ebenfalls zusammen mit den wasserlöslichen Enzymen zu mahlen.These additives, as well as the above. fat-soluble vitamins and carotenoids can be added before, during or after the grinding of the suspension according to the invention. In order to obtain a homogeneous suspension of all non-oil-soluble components that is as fine as possible, it is advantageous to use the above-mentioned Grind additives together with the water-soluble enzymes.
Als Mineralstoffe können beispielsweise Eisensulfat, Zinksulfat, Mangansulfat, Kupfersulfat, Calciumsulfat, Natriumsulfat, Kupfer- oxid, Magnesiumoxid, Calciumfluorid, Kaliumchlorid, Kaliumjodid, Natriumchlorid, Calciumjodat, Calcium-, Magnesium-, Kalium-, Natrium- oder Eisen-Phosphat, Cobaltcarbonat, Natriumselenat oder Kieselsäure und deren Salze in die Suspension eingearbeitet und mitvermahlen werden. Die Menge an eingesetzten Mineralstoffe , beispielsweise im Tierernährungsbereich, orientiert sich dabei jeweils am Bedarf der zu fütternden Tiere.As minerals, for example iron sulfate, zinc sulfate, manganese sulfate, copper sulfate, calcium sulfate, sodium sulfate, copper oxide, magnesium oxide, calcium fluoride, potassium chloride, potassium iodide, sodium chloride, calcium iodate, calcium, magnesium, potassium, sodium or iron phosphate, cobalt carbonate, Sodium selenate or silica and their salts are incorporated into the suspension and also ground. The amount of minerals used, for example in the animal nutrition sector, is based on the needs of the animals to be fed.
Als Aminosäurereste kommen generell alle bekannten physiologisch unbedenklichen α-Aminosäurereste in Frage. Bevorzugt zu nennen sind die Reste folgender Aminosäuren: Alanin, Arginin, Asparagin, Asparaginsäure, Cystein, Cystin, Glutamin, Glutaminsäure, Glycin, Histidin, Isoleucin, Leucin, Lysin, Methionin, Phenylalanin, Hippursäure, Serin und Taurin. Besonders bevorzugt sind Lysin, Methionin und Cystein.All known physiologically harmless α-amino acid residues are generally suitable as amino acid residues. The residues of the following amino acids are preferred: alanine, arginine, asparagine, aspartic acid, cysteine, cystine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, hippuric acid, serine and taurine. Lysine, methionine and cysteine are particularly preferred.
Weitere Bestandteile der Suspension können sein: Verbindungen mit Vitamin- oder Coenzymcharakter, z . B . Cholin- chlorid, Carnitin, γ-Butyrobetain, Liponsäure , Kreatin, Ubichinone, S-Methylmethionin, S-Adenosylmethionin.Other components of the suspension can be: Compounds with a vitamin or coenzyme character, e.g. B. Choline chloride, carnitine, γ-butyrobetaine, lipoic acid, creatine, ubiquinones, S-methyl methionine, S-adenosyl methionine.
Mehrfach ungesättigte Fettsäuren, z . B . Linolsäure, Linolensäure , Arachidonsäure, Eicosapentaensäure, Docosahexaensäure .Polyunsaturated fatty acids, e.g. B. Linoleic acid, linolenic acid, arachidonic acid, eicosapentaenoic acid, docosahexaenoic acid.
Fütterungsantibiotika für Medizinalfutter sowie Mikroorganismen zur Verbesserung der Verdauung.Feeding antibiotics for medicated feed and microorganisms to improve digestion.
In manchen Fällen kann es erforderlich sein, daß die öligen Suspensionen außerdem Hilfsstoffe, wie z.B. Schutzkolloide, Anti- oxidantien, Verdicker, Chelatbildner, wie z.B. Alkali- oder Erdalkalisalze der Citronensäure, Phytinsäure oder Phosphorsäure enthalten und/oder Emulgatoren enthalten.In some cases it may be necessary that the oily suspensions also contain adjuvants such as e.g. Protective colloids, antioxidants, thickeners, chelating agents, such as e.g. Contain alkali or alkaline earth salts of citric acid, phytic acid or phosphoric acid and / or contain emulsifiers.
Als Schutzkolloide können beispielsweise Gelatine, Fischgelatine, Stärke, Dextrin, Pflanzenproteine, Pektin, Gummi-Arabikum, Kasein, Kaseinat oder Mischungen davon verwendet. Es können aber auch Polyvinylalkohol, Polyvinylpyrrolidon, Methylcellulose, Carboxymethylcellulose, Hydroxypropylcellulose und Alginate eingesetzt werden. Bezüglich näherer Einzelheiten wird auf R.A. Morton, Fat Soluble Vitamins, Intern. Encyclopedia of Food and Nutrition, Bd.9, Pergamon Press 1970, S. 128-131, verwiesen.Protective colloids that can be used are, for example, gelatin, fish gelatin, starch, dextrin, plant proteins, pectin, gum arabic, casein, caseinate or mixtures thereof. However, polyvinyl alcohol, polyvinyl pyrrolidone, methyl cellulose, carboxymethyl cellulose, hydroxypropyl cellulose and alginates can also be used. For more details, see R.A. Morton, Fat Soluble Vitamins, Intern. Encyclopedia of Food and Nutrition, Vol. 9, Pergamon Press 1970, pp. 128-131.
Zur Erhöhung der Stabilität des Wirkstoffes gegen oxidativen Abbau ist es vorteilhaft, Stabilisatoren wie α-Tocopherol, t-Butyl-hydroxy-toluol, t-Butylhydroxyanisol, oder Ethoxyquin zuzusetzen.To increase the stability of the active substance against oxidative degradation, it is advantageous to add stabilizers such as α-tocopherol, t-butyl-hydroxy-toluene, t-butylhydroxyanisole or ethoxyquin.
Als Emulgatoren bzw. Solubilisatoren können beispielsweise Poly- glycerin-Fettsäureester , Sorbitan-Fettsäurees ter , Propylenglycol- Fettsäureester oder Lecithin verwendet werden.Examples of emulsifiers or solubilizers which can be used are polyglycerol fatty acid esters, sorbitan fatty acid esters, propylene glycol fatty acid esters or lecithin.
Die Suspensionen eignen sich u. a. als Zusatzstoff in Tierfuttermittelzubereitungen bzw. in Mischfutter und zur Herstellung von Nahrungsergänzungspräparaten im Tierbereich.The suspensions are u. a. as an additive in animal feed preparations or in compound feed and for the production of nutritional supplements in the animal sector.
Bevorzugt lassen sich die Suspensionen als Futtermittelzusatz in der Tierernährung einsetzen, bevorzugt zum Auftragen bzw. "Aufsprühen auf Futtermittelpellets , besonders bevorzugt zum Zumischen zu Futtermittelmischungen vor dem Pelletieren. Die Anwendung als Futtermittelzusatzstoff erfolgt insbesondere durch direktes Aufsprühen der erfindungsgemäßen Suspensionen, gegebenenfalls nach Verdünnung mit Ölen, beispielsweise auf Tierfutterpellets als sogenannte "post-pelleting-application" .The suspensions can preferably be used as a feed additive in animal nutrition, preferably for application or "spraying" onto feed pellets, particularly preferably for admixing with feed mixtures before pelleting. It is used as a feed additive in particular by spraying the suspensions according to the invention directly, optionally after dilution with oils, for example on animal feed pellets as a so-called "post-pelleting application".
Eine bevorzugte Ausführungsform des Sprühverfahrens besteht darin, daß man die Futtermittelpellets unter vermindertem Druck mit der öligen Suspension belädt.A preferred embodiment of the spraying process consists in loading the feed pellets with the oily suspension under reduced pressure.
Beispiele hierfür finden sich u.a. in GB-A-2 232 573 sowie in EP-A-0 556 883.Examples include: in GB-A-2 232 573 and in EP-A-0 556 883.
Bevorzugt richtet sich die Erfindung auf Tierfuttermittel, insbesondere auf Futtermittelpellets, die mit den Suspensionen beladen werden oder vor dem Pelletieren der Futtermittelmischungen in gewünschten Kombinationen zugemischt werden.The invention is preferably directed to animal feed, in particular feed pellets which are loaded with the suspensions or are mixed in desired combinations before the feed mixtures are pelletized.
Die nach dem erf ndungsgemäßen Verf hren herstellbaren Öligen Suspensionen weisen den Vorteil einer erhöhten Lagerstabilität auf und eine verminderte Anfälligkeit gegenüber mikrobiologischen Verkeimungen.The oily suspensions which can be produced by the process according to the invention have the advantage of increased storage stability and reduced susceptibility to microbiological contamination.
Zusätzlich können die öligen Suspensionen ohne den aufwendigen Verfah.renssch.ritt der Umhüllung direkt äen Puttermittelmischungen vor dem Pelletieren zugesetzt werden.In addition, the oily suspensions can be added directly to putter mixes prior to pelleting without the complex procedural step of the coating.
//
In den folgenden Beispielen wird die Herstellung der erfindungs- gemäßen öligen Suspensionen wasserlöslicher Enzyme näher erläutert .The preparation of the oily suspensions of water-soluble enzymes according to the invention is explained in more detail in the following examples.
Beispiel 1example 1
Zwei Kilogramm einer Mischung aus 25 Gew.-% getrocknete Phytase (aus Asperg-illus, sprühgetrocknet), und 75 Gew.-% Maiskeimöl werden mit einem Blattrührer solange gerührt, bis eine homogene Suspension vorliegt. Danach wird die Mischung in eine rührbare Vorlage umgefüllt, aus der die Suspension mittels Schlauchpumpe durch eine kontinuierlich betriebene Kugelmühle (Dyno Mil-TKDL Spezial) gefördert wird. Der Mahlbehälter der Kugelmühle ist mit 400 g Glaskugeln (Durchmesser 800 bis 1200 μm) gefüllt. Die aus der Mühle austretende feinteilige Suspension wird aufgefangen und mittels eines Partikelgrößenmeßgerätes (Malvern Mastersizer) vermessen. Der MahlVorgang wird so oft wiederholt bis die suspendierten Teilchen ein mittlere Teilchengröße kleiner 20 μm haben. Two kilograms of a mixture of 25% by weight of dried phytase (from Asperg-illus, spray-dried) and 75% by weight of corn oil are stirred with a blade stirrer until a homogeneous suspension is obtained. The mixture is then poured into a stirrable receiver, from which the suspension is conveyed by means of a peristaltic pump through a continuously operated ball mill (Dyno Mil-TKDL Special). The grinding bowl of the ball mill is filled with 400 g glass balls (diameter 800 to 1200 μm). The fine-particle suspension emerging from the mill is collected and measured using a particle size measuring device (Malvern Mastersizer). The grinding process is repeated until the suspended particles have an average particle size of less than 20 μm.

Claims

Patentansprüche claims
1. Verfahren zur Herstellung öliger Suspensionen wasserlöslicher Enzyme, dadurch gekennzeichnet, daß man in Abwesenheit wasserlöslicher Vitamine1. A process for the preparation of oily suspensions of water-soluble enzymes, characterized in that in the absence of water-soluble vitamins
a) mindestens ein wasserlösliches Enzym in einem Öl bis auf eine mittlere Partikelgröße von 0,1 bis 100 μ mahlt odera) grinding at least one water-soluble enzyme in an oil to an average particle size of 0.1 to 100 μ or
b) mindestens ein wasserlösliches Enzym ohne Verwendung einer kontinuierlichen Phase bis auf eine mittlere Partikelgröße von 0,1 bis 100 μm mahlt und die gemahlenen Partikel anschließend in einem Öl suspendiert .b) grinding at least one water-soluble enzyme without using a continuous phase down to an average particle size of 0.1 to 100 μm and then suspending the ground particles in an oil.
** * *
2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß es sich bei dem Öl um mindestens ein eßbares Öl handelt.2. The method according to claim 1, characterized in that the oil is at least one edible oil.
3. Verf hren nach einem der Ansprüche 1 oder 2 , dadurch gekenn- zeichnet, daß es sich bei dem Öl um ein bei 20°C flüssiges Öl handelt .3. Procedure according to one of claims 1 or 2, characterized in that the oil is an oil which is liquid at 20 ° C.
4. Verfahren nach einem der Ansprüche 2 oder 3 , dadurch gekennzeichnet, daß man als eßbares Öl Vitamin E, Vitamin E- Derivate oder Mischungen davon verwendet.4. The method according to any one of claims 2 or 3, characterized in that vitamin E, vitamin E derivatives or mixtures thereof are used as edible oil.
5. Verfahren nach einem der Ansprüche 1 bis 4 , dadurch gekennzeichnet, daß die Mahlung im Schritt a) sowie die Mahlung und/oder Suspendierung in Schritt b) in Abwesenheit eines Emulgators erfolgen.5. The method according to any one of claims 1 to 4, characterized in that the grinding in step a) and the grinding and / or suspension in step b) are carried out in the absence of an emulsifier.
6. Verfahren nach einem der Ansprüche 1 bis 5 , dadurch gekennzeichnet, daß die Mahlung im Schritt a) sowie die Mahlung und/oder Suspendierung in Schritt b) in Abwesenheit eines Schutzkolloids erfolgen.6. The method according to any one of claims 1 to 5, characterized in that the grinding in step a) and the grinding and / or suspension in step b) take place in the absence of a protective colloid.
7. Verfahren nach einem der Ansprüche 1 bis 6 , dadurch gekennzeichnet, daß man mindestens ein wasserlösliches Enzym als Feststoff mit einem Wassergehalt kleiner 10 Gew.-% verwendet.7. The method according to any one of claims 1 to 6, characterized in that at least one water-soluble enzyme is used as a solid with a water content of less than 10 wt .-%.
Verfahren nach einem der Ansprüche 1 bis 7 , dadurch gekennzeichnet, daß die Mahlung im Schritt a) sowie die Suspendierung in Schritt b) in Gegenwart fettlöslicher Vitamine erfolgen. Method according to one of claims 1 to 7, characterized in that the grinding in step a) and the suspension in step b) are carried out in the presence of fat-soluble vitamins.
9. Verfahren nach einem der Ansprüche 1 bis 8, dadurch gekennzeichnet, daß die Mahlung im Schritt a) sowie die Mahlung und/oder Suspendierung in Schritt b) in Gegenwart von Trok- kenmitteln erfolgen.9. The method according to any one of claims 1 to 8, characterized in that the grinding in step a) and the grinding and / or suspension in step b) are carried out in the presence of drying agents.
55
10. Verfahren nach Anspruch 9, dadurch gekennzeichnet, daß man als Trockenmitteln Stoffe, ausgewählt aus der Gruppe, bestehend aus Alkali- und Erdalkalisulfaten, Alkali- und Erdalkalichloriden und Kieselgel verwendet.10. The method according to claim 9, characterized in that substances selected from the group consisting of alkali and alkaline earth metal sulfates, alkali metal and alkaline earth metal chlorides and silica gel are used as drying agents.
1010
11. Ölige Suspensionen mindestens eines wasserlöslichen Enzyms, erhältlich nach einem Verfahren, definiert gemäß einem der Ansprüche 1 bis 10.11. Oily suspensions of at least one water-soluble enzyme, obtainable by a process defined according to one of claims 1 to 10.
15 12. Ölige _Suspensionen nach Anspruch 11, enthaltend 1 bis 70 Gew.-% mindestens eines wasserlöslichen Enzyms.15 12. Oily _Suspensions according to claim 11, containing 1 to 70 wt .-% of at least one water-soluble enzyme.
13. Ölige Suspensionen nach einem der Ansprüche 11 oder 12 , enthaltend zusätzlich 0,5 bis 60 Gew.-% mindestens eines13. Oily suspensions according to one of claims 11 or 12, additionally containing 0.5 to 60% by weight of at least one
20 fettlöslichen Vitamins.20 fat-soluble vitamins.
14. Verwendung der öligen Suspensionen, definiert gemäß einem der Ansprüche 11 bis 13 als Zusatz zu Tierfuttermitteln.14. Use of the oily suspensions, defined according to one of claims 11 to 13, as an additive to animal feed.
25 15. Verwendung nach Anspruch 14 als Futtermittelzusatz in der Tierernährung .25 15. Use according to claim 14 as a feed additive in animal nutrition.
16. Verwendung nach Anspruch 15 zum Auftragen auf Futtermittelpellets .16. Use according to claim 15 for application to feed pellets.
3030
17. Verwendung nach Anspruch 15 zum Zumischen zu Futtermittelmischungen vor dem Pelletieren.17. Use according to claim 15 for admixing to feed mixtures before pelleting.
18. Verwendung nach Anspruch 16, dadurch gekennzeichnet, daß man 35 die Futtermittelpellets unter vermindertem Druck mit der öligen Suspension belädt.18. Use according to claim 16, characterized in that 35 the feed pellets are loaded with the oily suspension under reduced pressure.
19. Futtermittelpellets, enthaltend ölige Suspensionen, definiert gemäß einem der Ansprüche 11 bis 13.19. Feed pellets containing oily suspensions, defined according to one of claims 11 to 13.
40 """ 40 """
45 45
EP02777104A 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes Withdrawn EP1432795A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP06120633A EP1746155A1 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
DE10147035A DE10147035A1 (en) 2001-09-25 2001-09-25 Process for the preparation of oily suspensions of water-soluble enzymes
DE10147035 2001-09-25
PCT/EP2002/010321 WO2003029452A2 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes

Related Child Applications (1)

Application Number Title Priority Date Filing Date
EP06120633A Division EP1746155A1 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes

Publications (1)

Publication Number Publication Date
EP1432795A2 true EP1432795A2 (en) 2004-06-30

Family

ID=7700101

Family Applications (2)

Application Number Title Priority Date Filing Date
EP06120633A Withdrawn EP1746155A1 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes
EP02777104A Withdrawn EP1432795A2 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP06120633A Withdrawn EP1746155A1 (en) 2001-09-25 2002-09-14 Method for the production of oily suspensions of water-soluble enzymes

Country Status (8)

Country Link
US (1) US20040241285A1 (en)
EP (2) EP1746155A1 (en)
JP (1) JP2005505277A (en)
CN (1) CN1316017C (en)
CA (1) CA2460731A1 (en)
DE (1) DE10147035A1 (en)
MX (1) MXPA04002041A (en)
WO (1) WO2003029452A2 (en)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7416869B2 (en) * 2004-08-19 2008-08-26 Lonza Ltd. Enzyme delivery systems, application in water based products
JP2007519408A (en) * 2004-01-30 2007-07-19 ビーエーエスエフ　アクチェンゲゼルシャフト Stabilized enzyme preparation
DE602005007182D1 (en) * 2004-01-30 2008-07-10 Basf Se STABILIZED PHYTASE-CONTAINING GRANULES
AU2011215669B2 (en) 2010-02-12 2015-06-04 HedoniaUSA, Inc. Compositions and methods for treating depression
WO2014025203A1 (en) * 2012-08-08 2014-02-13 주식회사 씨티씨바이오 Water-soluble mannanase composition, and method for preparing same
CN103549160A (en) * 2013-11-01 2014-02-05 广州市博仕奥生物科技有限公司 Post-injection enzyme of puffed feed as well as preparation method and application of post-injection enzyme
WO2018144088A1 (en) 2016-11-03 2018-08-09 Alexander Vuckovic, M.D., Llc Compositions and methods for treating depression
CN107874070A (en) * 2017-11-08 2018-04-06 南京喜之郎食品有限公司 A kind of beet red pigment emulsifying process
DE102021131457A1 (en) 2021-11-30 2023-06-01 Farmerscent GmbH Crude fiber crust pellets, animal feed containing them and corresponding processes and uses

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NO158919C (en) * 1983-02-17 1988-11-16 Kibun Kk METHOD AND MEASURING MEASURES FOR MEAT MEAT.
EP0556883B1 (en) * 1992-01-24 1998-07-22 Gist-Brocades N.V. Method for the preparation of feed pellets
EP0896618B1 (en) * 1996-04-29 2007-06-20 Novozymes A/S Non-aqueous, liquid, enzyme-containing compositions
JPH09322770A (en) * 1996-05-31 1997-12-16 Ichibiki Kk Exogenous enzyme composition stably dispersed in oil and its production and usage
CN1197483C (en) * 2000-03-17 2005-04-20 巴斯福股份公司 Method of producing oily suspensions of water-soluble vitamins

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO03029452A2 *

Also Published As

Publication number Publication date
CA2460731A1 (en) 2003-04-10
WO2003029452A3 (en) 2003-09-12
US20040241285A1 (en) 2004-12-02
EP1746155A1 (en) 2007-01-24
DE10147035A1 (en) 2003-04-24
CN1558948A (en) 2004-12-29
MXPA04002041A (en) 2004-06-07
JP2005505277A (en) 2005-02-24
WO2003029452A2 (en) 2003-04-10
CN1316017C (en) 2007-05-16

Similar Documents

Publication Publication Date Title
EP1272059B1 (en) Method of producing oily suspensions of water-soluble vitamins
EP1928260B1 (en) Phytase-containing enzyme granulate ii
US20060078598A1 (en) Animal feed containing an encapsulated ingredient
EP0982038B1 (en) Stable powdery vitamin and carotenoide containing compositions and process to prepare them
AU2005279201B2 (en) Method for pigment solubilisation, a pigment composition and its use
EP0326026A2 (en) Stable mixture containing oxidation-sensitive compounds, process to obtain it and use of a combination of products to stabilize oxidation-sensitive compounds
EP2403362B1 (en) Powdered compositions of astaxanthin derivatives ii
WO2008087140A2 (en) Liquid formulations containing a carotinoid
EP1432795A2 (en) Method for the production of oily suspensions of water-soluble enzymes
EP2124615A1 (en) Unctuous compositions
EP2403364B1 (en) Method for producing solutions of astaxanthin derivatives
EP2403354B1 (en) Formulation of astaxanthin derivatives and use thereof
JP2000106844A (en) Water-dispersible carotenoid composition
EP1186292B1 (en) Dry carotinoid powder with a mulit-core structure
WO2004012707A1 (en) Ascorbic acid salt suspensions and use thereof as antioxidants
WO2010100227A1 (en) Formulation of astaxanthin derivatives and use thereof ii
DE10049137A1 (en) Production of oily suspensions of water soluble vitamins comprises milling of vitamins in oil or dry milling and suspension in oil
DE10013312A1 (en) Production of oily suspensions of water soluble vitamins comprises milling of vitamins in oil or dry milling and suspension in oil
RU2227513C1 (en) Method for preparing combined fodder premix
RU2227514C1 (en) Method for preparing combined fodder premix
RU2227509C1 (en) Method for preparing combined fodder premix
RU2226843C1 (en) Process of producing mixed fodder premix
RU2227512C1 (en) Method for preparing combined fodder premix
DE102008058449A1 (en) Producing carotenoid-solutions in edible oil, comprises providing a carotenoid suspension in oil, heating a stream of the suspension in edible oil to produce a hot carotenoid-solution and cooling the hot carotenoid-solution

Legal Events

Date Code Title Description
PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

Free format text: ORIGINAL CODE: 0009012

17P Request for examination filed

Effective date: 20040426

AK Designated contracting states

Kind code of ref document: A2

Designated state(s): AT BE BG CH CY CZ DE DK EE ES FI FR GB GR IE IT LI LU MC NL PT SE SK TR

AX Request for extension of the european patent

Extension state: AL LT LV MK RO SI

RIN1 Information on inventor provided before grant (corrected)

Inventor name: HEINZL, WOLFGANG

Inventor name: SOERENSEN, PREBEN

Inventor name: BRAUN, JOERG

Inventor name: RUNGE, FRANK

Inventor name: HABICH, ANDREAS

RAP1 Party data changed (applicant data changed or rights of an application transferred)

Owner name: BASF SE

STAA Information on the status of an ep patent application or granted ep patent

Free format text: STATUS: THE APPLICATION IS DEEMED TO BE WITHDRAWN

18D Application deemed to be withdrawn

Effective date: 20080429