CN1201372A - Feed enzyme products - Google Patents
Feed enzyme products Download PDFInfo
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- CN1201372A CN1201372A CN96197980A CN96197980A CN1201372A CN 1201372 A CN1201372 A CN 1201372A CN 96197980 A CN96197980 A CN 96197980A CN 96197980 A CN96197980 A CN 96197980A CN 1201372 A CN1201372 A CN 1201372A
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- enzyme
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- 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
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- 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
- A23K40/10—Shaping or working-up of animal feeding-stuffs by agglomeration; by granulation, e.g. making powders
-
- 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
- A23K40/25—Shaping or working-up of animal feeding-stuffs by extrusion
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Polymers & Plastics (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Animal Husbandry (AREA)
- Fodder In General (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
A particulate, enzyme-containing preparation suitable for use, e.g., in the manufacture of an animal feed composition comprises: a total of at least 1 % by weight (w/w) of one or more hydrophobic substances; and a total of at least 75 % (w/w) of one or more water-insoluble substances, including the hydrophobic substance(s).
Description
The present invention relates to the granular enzyme preparation (as the goods (hereinafter often abbreviating " particle " as) of particle form) that contains, when described enzyme is applicable to composition as animal-feed, it is specially adapted to mix in the animal feedstuff compositions, particularly is mixed with the animal feedstuff compositions or the analogue of piller (" Pelleting ").When using, during as the steam pill, the invention enables the high composition of production enzymic activity reserving degree to become possibility at the industrial working condition that often is preferred for this based composition.
Proved the enzyme with some type, beta-glucanase for example, zytase or phytase mix in the animal feedstuff compositions can relevant animal absorbs nutrition from feed or mineral substance has obvious favorable influence to improving.In fodder production industries,, comprise being easy to handle with quantitative owing to the animal feedstuff compositions that various reasons are preferably prepared with the piller form; Low-level powder forms and is easy to be digested by animal during the processing.
Another important reasons of preferred pellets preparation is that pelletizing process itself helps feed is heat-treated, so that as removing invasive organism, belong to kind, campylobacter kind etc. as salmonella kind, listeria spp; For example, in order effectively to remove the salmonella kind, feed must thermal treatment under about at least 81 ℃ temperature in pelletizing process.In pelletizing process by means of as the steam treatment of feed is highly suitable for realizing this thermal treatment.
Although the thermal treatment to feed is important for above-mentioned reasons, but clearly should handle, and particularly relate to the processing that makes the feed composition be exposed to the use water vapor in heat and the steam and generally can reckon with for the enzyme that is present in the pending said composition to have harmful (deactivation or sex change) influence.Mention following document in this by way of example:
(I) Novo Nordisk publication A-06293 (can be from Denmark Novo Nordisk A/S, Bagsvaerd, obtain), the data of enzyme stability when it has provided the composition pill, wherein enzyme is respectively zytase and the beta-glucanase that contains from Humicola insolens, from the amylase of subtilis and from the bag of the proteolytic enzyme of Bacillus licheniformis by the particulate form.It is reported that in each case steam heating to 85 ℃ causes losing about 25% initial enzymic activity.
(ii) Novo Nordisk catalog number B402e-GB (can be from Denmark Novo NordiskA/S, Bagsvaerd, obtain), it has been reported and has been called Bio-Feed
TMPlus CT (contains the carbohydrase goods (Bio-Feed through Humicola insolens submergence fermentative production
TMThe enzyme granulate of bag quilt Plus)) is heated to (comprising this enzyme granulate) to keep in 83 ℃ the feed pelletizing process at the feed composition and surpasses 75% enzymic activity.
(iii) EP 0569468 B1 discloses (seeing wherein embodiment 1) and wherein is called " Bio-FeedPlus T " (latter is so-called " T-particle " type particle, promptly, particle according to US4106991 production, except enzyme, it comprises the cellulosic fibre of the meticulous fracture of 2-40%w/w especially) the bag of particulate state enzyme preparation mixed as the composition of feed composition by form and it related to 70 ℃ of temperature and open steam inject and when continuing the pill program of 25-30 second, keep the original fungi 1,4 beta-glucanase activity of its 90-100%.On the contrary, " Bio-Feed Plus " particle that does not wrap the T-particle of quilt and do not belong to the corpuscular bag quilt of T-all shows the fungi 1,4 beta-glucanase activity of reservation lower (75%).
People know different enzymes can have extensively different feature, and particularly for its thermostability, for example it tolerates the ability of quite high temperature.And, to be bag can be shown extensively different stability (thermostability particularly by T-granule type enzyme containing granule (on seeing) according to the characteristic of non-enzyme component in this particle and content to the inventor's experience, for example, stability in the pelletizing process), also be like this for bag by the enzyme containing granule beyond the T-granule type.
Therefore basic goal of the present invention be obtain particular form the used course of processing in the animal-feed manufacture (for example, pelletizing process) (under those conditions of simply listing in the above) have the zymin of high stability, for example enzyme containing granule under the heating and steam condition in.
The inventor is surprised to find process at granular (i) hydrophobic substance that comprises suitable proportion in the agent prepared from enzyme that contains, particularly work as some or all hydrophobic substances (for instance, it can be the wax material or such as the fatty substance of tallow) when being present in the bag tegillum that enwrapped granule contains the enzyme core, (ii) water-fast material (comprising above-mentioned hydrophobic substance) can be implemented in obviously increases the reservation that particulate state contains the enzymic activity of enzyme preparation (for example, particle) under the above-mentioned pill condition.
Therefore, a first aspect of the present invention relates to a kind of granular enzyme preparation (for example, a kind of particle) that contains, and comprises one or more hydrophobic substances that account at least 1% weight (w/w) altogether; Reach and account for one or more water-fast materials that comprise this hydrophobic substance of 75%w/w at least altogether.
Preferred hydrophobic substance constitutes the 5%w/w at least that contains enzyme preparation, more preferably about at least 8%w/w.Water-fast material preferably constitutes 80%w/w at least, more preferably 85%w/w at least, for example at least 90% contain enzyme preparation.
Described weight percentage is based on finished product gross weight (that is, comprising any bag tegillum that for example may exist).
Do not wrap by goods of the present invention, general preferred hydrophobic substance is distributed in the material of product grain (or be distributed at least in the typical particulate matter of taking from goods) basically equably.The suitable content of hydrophobic substance is in the scope of 5-95%w/w in these goods.In some cases, in addition the content of high hydrophobic substance to about 98%w/w also can be suitable for this class particulate state and contain enzyme preparation.
As mentioned above, preferred particulate state of the present invention contains enzyme preparation and comprises that (a) contains the enzyme core, (b) contains hydrophobic substance and surrounds or wrap up the bag tegillum that contains the enzyme core.In these goods, hydrophobic substance constitutes 1 to 50%w/w of goods, usually is from 5 to 25%w/w, more typically is 5 to 15%w/w.
Contain the particle that term used in the explanation of enzyme preparation " particle " refers to comprise Any shape or form of the present invention; as has certain particle of geometrical shape of rule (as sphere) basically; erose particle, or be the rule or the particle of irregular sheet form and have different shapes or the granular mixture of form (as the mixture of the grain type hereinafter mentioned).
Enzyme:
The enzyme classification of mentioning in this specification and claims book number (EC number) is according to the recommendation (1992) of biochemical and NK of molecular biology international association, academic publishing company, 1992.
The enzyme that the present invention contains in the enzyme preparation can be any type of enzyme, particularly relevant with animal feedstuff compositions any type of enzyme.Therefore, for example, this enzyme can be selected from phytase (3-phytase (that is inositol-six phosphoric acid 3-phosphohydrolase, particularly; Be categorized as EC3.1.3.8) or 6-phytase (that is inositol-six phosphoric acid 6-phosphohydrolase; Be categorized as EC3.1.3.26)), Phosphoric acid esterase (for example, be categorized as the enzyme of EC3.1.3.1 or EC3.1.3.2), zytase (for example, be categorized as EC3.2.1.8 or EC3.2.1.32 enzyme), beta-glucanase (for example, be categorized as EC3.2.1.75, EC3.2.1.71, the enzyme of EC3.2.1.59. or EC3.2.1.39), the paragalactan enzyme (for example, be categorized as the enzyme of EC3.2.1.89 (1.4-paragalactan restriction endonuclease) or EC3.2.1.90 (1,3-paragalactan restriction endonuclease)), alpha-galactosidase is (for example, be categorized as the enzyme of EC3.2.1.22), beta-galactosidase enzymes (for example, being categorized as the enzyme (Sumylact L) of EC3.2.1.23), α-Dian Fenmei (EC3.1.1.1), beta-amylase (EC3.2.1.2), cellulase (for example, being categorized as the enzyme of EC3.2.1.4), polygalacturonase (being categorized as the enzyme of EC3.2.1.15) and peptase (EC3.4), comprise " protein enzyme " or " proteolytic enzyme ", as be categorized as the enzyme of EC3.4.21.
Clear from top description, can the enzyme that one or more are different mix particulate state of the present invention and contain in the enzyme preparation.The enzyme composition that contains in the enzyme preparation particle of the present invention is seemingly unessential in intragranular distribution.Therefore, this enzyme composition can be evenly or is distributed in unevenly in the given particulate matter.
If the particle of bag quilt, particulate enzyme composition is confined to the particulate core basically, but will be appreciated that at least some enzyme compositions can be present in the bag tegillum.
The zymoprotein content general range that contains in the enzyme preparation in particulate state of the present invention is a 0.01-10% weight, as accounts for the 0.1-10% heavy (w/w) of goods, more typically is 0.2-5%w/w, normally 0.3-2.5%w/w.Enzyme content depends on the enzymatic property that is present in the particulate state goods largely.The particulate state goods also can contain other material from the enzyme production process, are that the general enzyme preparation of incomplete purifying that adopts is used for mixing particle product in industrial embodiment preferred.
Particle:
As (the seeing above) that illustrated to a certain extent, it is particle (that is, granulate form) that particulate state of the present invention contains that enzyme preparation is very suitable for, as the particle of " T-particle " (seeing above) or some other types.Perhaps, the particle that particulate state of the present invention contains enzyme preparation can be as containing the particle as the fusible composition of the matrix of enzyme and other composition of suitable proportion, fusible composition for example is plam oil (and/or other meltability vegetables oil or fat), hydrogenated palm kernel oil (and/or other hydrogenated vegetable oil), tallow, hydrogenated tallow or wax.In this class particulate was produced, the relevant composition with other of the enzyme of these goods imported in the fusible composition that melts, and this thawing thing is solidified.
Contain particulate size in the enzyme preparation as for particulate state of the present invention, consider that in order to help with other composition thorough mixing of feed and to guarantee that enzyme distributes enough evenly in the final feed product, the feasible upper limit of waiting to mix most of particulate granular sizes of the goods in the feed composition generally is about 2mm.
In this manual, the big I of particulate is regarded particulate max line molded dimension as, and therefore, for example, if be spheric particle (as being the spheric granular particle basically) basically, described granular size can be this particulate diameter.
The lower limit that contains the enzyme preparation granular size of the present invention general main by in the production of these goods and the needs of avoiding forming dust during handling determine.The feasible lower limit of this granular size usually is about 0.1mm.
Hydrophobic substance:
Term " hydrophobic substance " used in the specification sheets of the present invention refers to be not easy by water-moistened material, that is, it tends to repel water.Water insoluble fully in this material (its example is an oil, fat, the resin of chloroflo and many types) general essence.
Relevant especially in this manual hydrophobic substance generally is the material that dissolves in hydrocarbon organic solvent (for example, hexane, heptane etc.) or chlorinated hydrocarbon organic solvent (for example, methylene dichloride, chloroform etc.).Its suitable example comprises various glyceryl ester (that is, monoglyceride, diester or three esters), as Tallow, beef (for example, tallow or suet) and vegetables oil and some derivative thereof.
The hydrophobic substance of particularly suitable is that those are at ambient temperature for solid and have the material of about 40 ℃ or above fusing point.Its example comprises natural or sclerosis (hydrogenation) vegetables oil or fat such as some, for example, and hydrogenated palm kernel oil, the material of hydrogenated palm kernel oil or hydrogenated soybean oil and such as the material of hydrogenated tallow (for example, hydrogenated tallow or suet).
Generally being suitable for measuring particulate state of the present invention contains hydrophobic substance (as defined herein) total content in the enzyme preparation and utilizes methylene dichloride and the mensuration program of normal hexane extraction is partly described (seeing below) at this paper material and method.
Water-fast material
In the specification sheets of the present invention used term " water-fast material " comprise from basically fully water-fast those materials to those water, show quite low solvability (as, said solubleness is that every 100ml pure water is approximately 1g at most) the material scope.Therefore, water-fast material includes but not limited to the hydrophobic substance that is defined as above in specification sheets of the present invention.Other the type water-fast material relevant with the present invention (for example comprises various inorganic salt, lime carbonate, calcium sulfate, secondary calcium phosphate, magnesiumcarbonate), comprise that pure aluminium silicate (for example, clay,, bentonite or Fuller's earth as kaolin) mineral substance, inert metal oxides (for example, titanium dioxide or magnesium oxide), various macromolecular substance (for example, polysaccharide, starch as Mierocrystalline cellulose or some type, and levigated cereal or cereal powder (for example, from such as wheat, barley, the cereal production of rye) or soybean (for example, rough powder of soybean or soya flour)) and activated carbon.
Generally be suitable for measuring particulate state according to the present invention and contain the mensuration program of water-fast material (as defined herein) total content in the enzyme preparation in this paper material and method part (seeing below) description.
Others of the present invention:
The invention still further relates to the method that contains the enzyme animal feedstuff compositions of producing, wherein to carrying out steam heating step (as the pill program that is usually directed to list above, wherein the final feed composition becomes ball) with other composition blended of final feed composition enzyme preparation that contains of the present invention.
Another aspect of the present invention relates to particulate state of the present invention and contains the application of enzyme preparation in the production that contains the enzyme animal feedstuff compositions.
Relevant therewith, the present invention also comprises: use particulate state of the present invention to contain enzyme preparation as containing the enzyme-containing feed composition that the enzyme initial substance is produced; And with (above list) obtain or obtainable enzyme animal feedstuff compositions that contains in accordance with the present production process.
It is evident that the enzyme containing granule shape goods of preparing according to the present invention also can be used for its production needs heat treated other (non-feed) as herein described and contain in the production of enzyme composition.
Can imagine that further principle of the present invention not only can be applicable to improve and (for example contain enzyme preparation thermal treatment, the reservation of the enzymic activity that steam treatment) interrelates, and can be used for improving VITAMIN similarly, assist VITAMIN, amino acid, medicine (for example, microbiotic or growth regulatory substance), the stability of inorganic additive etc. or the vigor of raising microorganism (for example, bacterium or fungi), promptly after the particle product of these materials is mixed in thermal treatment (for example, steam treatment).
The present invention has carried out further instruction with the embodiment that material and method below partly provide, but is not to limit on any degree.
Material and method: the general description of T particle product
As described in the past, " T particle " type particle has carried out feature description in the U.S. 4,106,991.
Below the bag of describing among the embodiment 1 and 2 by the T-particle (they all are particulate examples, wherein required hydrophobic substance mix bag by in) initial substance below adopting prepares with described general method hereinafter:
1) cellulosic fibre (Arbocel
TMBC200),
2) levigated " weighting agent " (for goods of the present invention, it comprises one or more water-fast materials of the present invention),
3) dextrin (TACKIDEX
TMGM155 or AVIDEX
TM28LA21) and/or another kind of carbohydrate wedding agent, and
4) aqueous solution of enzyme.(further details reference embodiment 1 and 2 (seeing below)).
In being equipped with the Lodige mixing tank of essential mixed pulp and rotating knife, granulate.
After cellulosic fibre and weighting agent and part wedding agent are thoroughly mixed, will contain enzyme aqueous solution and remain wedding agent and spray in the mixing tank, the effect by rotating knife causes forming granular particle.
To carry out drying in the transfer of granules influent stream movable bed moisture eliminator then, afterwards with dry granules cooling and sieve.
Also in the Lodige mixing tank, carried out by gained T granular particles with the dressing bag that contains hydrophobic substance.
The exsiccant granular particle is heated to temperature in about 70-85 ℃ scope, normally 75-85 ℃ (the fusing feature that depends on used hydrophobic composition).Melt hydrophobic composition and in mixing tank, evenly be sprayed onto on the granular particle.Mix second weighting agent in the described mode of EP 0569468 B1.Behind using hydrophobic composition and the weighting agent, the T type particle of gained is cooled to envrionment temperature and sieves to remove fines and unwanted large-scale particle.
The main granular sizes of all T particles (diameter) that adopt among the embodiment that provides hereinafter are the 0.3-1.2mm scope.Measure the test of hydrophobic substance total content in the goods of the present invention:
20-25 ℃ of intermittent type in the 50ml methylene dichloride of about 10 gram described goods (accurate weight W gram) in the container of deadend stirred about 4 hours, the normal hexane that adds the 50ml equal portions, and then sealed vessel, stirring and make it to leave standstill several hours, liquid phase is limpid up to top.10ml is clarified the liquid phase sample transfer in the glass beaker of 50ml known weight (to 4 decimals), solvent is evaporated fully.And then weighing beaker (also to 4 decimals) is to measure the weight (R is a unit with the gram) of evaporation back residue.
The total content percentage ratio of hydrophobic materials is calculated as ((R * 10)/W) * 100 then.Measure the test of water-fast material total content in the goods of the present invention
The approximately described goods of 10g (accurate weight W gram) 20-25 ℃ of stirring 2 hours in the 50ml of encloses container deionized water.Centrifugal gained suspension/slurry samples, employing ° Brix measure through the refractive power art and measure the percentage ratio S that does solids content in the clarification supernatant, are accurate to 0.1%.
So water-fast material total content percentage ratio is calculated as: 100-((S * 50)/W).Embodiment 1 contains the preparation of the granular product of phytase: prepared 5 kinds of different particles that contain phytase with T-granulating program (seeing above).Phytase solution used in the production of phytase solution that uses in this granulating and product phytase Novo L (see Novo Nordisk products catalogue B-722a, can be from Denmark Novo NordiskA/S, Bagsvaerd asks for) is from identical source.Phytase Novo L is the liquid agent prepared from enzyme with stdn phytase activity.
Use the following component preparation difference called after PA of institute's limited amount (providing as the basis) with dry-matter, PB, PC, 5 kinds of particles of PD and PE: particle PA (the bag quilt) component accounts for the %w/w of goods
Core dressing sodium-chlor 54 kaolin 39 cellulosic fibres 9 lime carbonate 9 dextrin+sucrose+enzyme 8 hydrogenated tallows 9
It is evident that from last table: used main weighting agent is a water-soluble salt sodium-chlor among particle PA, and hydrophobic substance (hydrogenated tallow) and water-fast material (hydrogenated tallow+kaolin+Mierocrystalline cellulose+lime carbonate) total content is respectively 9%w/w and 39%w/w.The value of this value and employing said determination program determination is identical (being respectively 8%w/w and 37%) very.The phytase activity that uses NovoNordisk analytical method KALSM-0403.01/01 (can be from Denmark Novo Nordisk A/S, Bagsvaerd asks for) to measure granular PA is 2400FYT/g.
Particle PB (not wrapping quilt) component accounts for the %w/w cellulosic fibre 11 lime carbonate 73 dextrin+enzyme 16 of goods
It is evident that from this table the main weighting agent that uses among the particle PB is water-fast salt lime carbonate, water-fast material total content is 84%w/w.This value is coincide in good condition with adopting the above-mentioned value (84%w/w) that is used for the mensuration program determination of water-fast material.This matches with the hydrophobic substance<1%w/w that uses the above-mentioned assay method that is used for hydrophobic substance to find not use hydrophobic substance (as defined herein).The phytase activity that uses Novo Nordisk analytical method KAL-SM-0403.01/01 (seeing above) to measure particle PB is 1760FYT/g.
Particle PC (the bag quilt) component accounts for the %w/w of goods
Core dressing kaolin 7 cellulosic fibres 6 lime carbonate 54 7 dextrin+enzyme 12 hydrogenated tallows 11
It is evident that from this table the main weighting agent that uses in particle PC is water-fast salt lime carbonate, hydrophobic substance (hydrogenated tallow) and water-fast material (hydrogenated tallow+kaolin+Mierocrystalline cellulose+lime carbonate) are respectively 11%w/w and 87%w/w.This value is very identical with the value (being respectively 10%w/w and 87%) that adopts mensuration program determination recited above.The phytase activity that uses Novo Nordisk analytical method KAL-SM-0403.01/01 (seeing above) to measure particle PC is 950FYT/g.Particle PD (the bag quilt) component accounts for the %w/w of goods
Core dressing kaolin 6.3 7.8 cellulosic fibres 9.4 lime carbonate 7.8 secondary calcium phosphates 48.5 dextrin+enzyme 14.5 hydrogenated palm kernel oils 5.5
It is evident that from this table used main weighting agent is water-fast salt secondary calcium phosphate (CaHPO in particle PD
4), the total content of hydrophobic substance (hydrogenated palm kernel oil) and water-fast material (hydrogenated palm kernel oil+kaolin+Mierocrystalline cellulose+lime carbonate+secondary calcium phosphate) is respectively 5.5%w/w and 79.8%w/w.The phytase activity that uses Novo Nordisk analytical method KAL-SM-0403.01/01 (seeing above) to measure particle PD is 1430FYT/g.Particle PE (the bag quilt) component accounts for the %w/w of goods
Core dressing kaolin 5.1 9.3 cellulosic fibre 7.9 lime carbonate, 9.3 sodium sulfate (anhydrous) 42.8 dextrin+enzyme 17.1 hydrogenated palm kernel oils 8.5
It is evident that from this table, the main weighting agent that adopts in particle PE is a water-soluble salt sodium sulfate, and the total content of hydrophobic substance (hydrogenated palm kernel oil) and water-fast material (hydrogenated palm kernel oil+kaolin+Mierocrystalline cellulose+lime carbonate) is respectively 8.5%w/w and 40.1%w/w.The phytase activity that uses Novo Nordisk analytical method KAL-SM-0403.01/01 (seeing above) to record particle PE is 2400FYT/g.Embodiment 2 contains the preparation of zytase particle product: prepared 5 kinds of different particles that contain zytase with T-granulating program (seeing above).The zytase solution that in this particle, uses with at product B io-Feed
TMWheat L (referring to, Novo Nordisk publication B-854a for example can be from Denmark Novo Nordisk A/S, Bagsvaerd asks for) production in used enzyme from identical source.Bio-Feed
TMWheat L is the liquid enzyme preparation with stdn xylanase activity.
Use following component preparation difference called after XA with institute's limited amount (providing as the basis) with dry-matter, XB, XC, 5 kinds of particles of XD and XE:
Particle XA (the bag quilt) component accounts for the %w/w of goods
Core dressing sodium sulfate (anhydrous) 55 cellulosic fibres 7 lime carbonate 3 18 dextrin+enzyme 10 hydrogenated tallows 8
It is evident that from last table used main weighting agent is a water-soluble salt sodium sulfate in particle XA, the total content of hydrophobic substance (hydrogenated tallow) and water-fast material (hydrogenated tallow+Mierocrystalline cellulose+lime carbonate) is respectively 8%w/w and 36%w/w.This value is very identical with the value (being respectively 9%w/w and 36%) that adopts mensuration program determination mentioned above.The xylanase activity that uses NovoNordisk analytical method KAL-SM-0356.01/01 (can be from Denmark Novo Nordisk A/S, Bagsvaerd asks for) to measure particle XA is 810FXU/g.Particle XB (the bag quilt) component accounts for the %w/w of goods
Core dressing calcium sulfate 54 kaolin 3 cellulosic fibres 7 lime carbonate 18 dextrin+enzyme 9 hydrogenated tallows 8
It is evident that from this table, used main weighting agent is water-fast salt calcium sulfate in particle XB, and hydrophobic materials (hydrogenated tallow) and water-fast material (hydrogenated tallow+calcium sulfate+kaolin+Mierocrystalline cellulose+lime carbonate) total content is respectively 8%w/w and 90%w/w.This value fits like a glove with the value (being respectively 8%w/w and 90%) that adopts the said determination program determination.The xylanase activity that uses Novo Nordisk analytical method KAL-SM-0356.01/01 (seeing above) to measure particle XB is 770FXU/g.Particle XC (not wrapping quilt) component accounts for %w/w sodium sulfate (anhydrous) the 76 cellulosic fibres 11 lime carbonate 6 dextrin+enzyme 8 of goods
It is evident that from this table used main weighting agent is a water-soluble salt sodium sulfate among the particle XC, water-fast material total content is 17%w/w.The described value that is used for the mensuration program determination of water-fast material is 23%w/w above adopting.Do not use hydrophobic substance (as defined herein), this is consistent with the hydrophobic substance<1%w/w that uses the above-mentioned mensuration that is used for hydrophobic substance to find.The xylanase activity that uses Novo Nordisk analytical method KAL-SM-0356.01/01 (seeing above) to measure among the particle XC is 1080FXU/g.Particle XD (not wrapping quilt) component accounts for the %w/w calcium sulfate 73 kaolin 4 cellulosic fibres 10 dextrin+enzyme 12 of goods
It is evident that from this table, used main weighting agent is water-fast salt calcium sulfate in particle XD, and the total content of water-fast material is that the value (88%w/w) of 87%w/w. this and the top described mensuration program determination that is used for water-fast material is coincide preferably.Do not use hydrophobic substance (as defined herein), this is consistent with hydrophobic substance<1%w/w that the above-mentioned mensuration that is used for hydrophobic substance is found.The xylanase activity that uses Novo Nordisk analytical procedure KAL-SM-0356.01/01 (seeing above) to measure particle XD is 1090FXU/g.Particle XE (not wrapping quilt) component accounts for the %w/w of goods
Core dressing cellulosic fibre 8 lime carbonate 54 18 dextrin+enzyme 12 hydrogenated tallows 8
It is evident that from last table used main weighting agent is water-fast salt lime carbonate in particle XE, hydrophobic substance (hydrogenated tallow) and water-fast material (hydrogenated tallow+Mierocrystalline cellulose+lime carbonate) total content is respectively 8%w/w and 88%w/w.This value is quite identical with the value (being respectively 6%w/w and 90%w/w) that adopts mensuration program determination recited above.The xylanase activity that uses Novo Nordisk analytical method KAL-SM-0356.01/01 (seeing above) to measure particle XE is 410FXU/g.
Embodiment 3
Steam heating is to the influence of the phytase activity of the feed preparation that contains phytase: checks to have mixed respectively to contain phytase particle product PA, and PB, PC, the reserving degree of enzymic activity after the feed preparation steam heating of one of PD and PE (seeing embodiment 1):
Select the cavings feed preparation of following composition (w/w):
62% wheat
15% soyflour
12% barley
7% fish meal
2% soybean oil
1% lime carbonate
1% Lin Suanergai
0.2%?Solivit
TM?Mikro?106
*
*Commercial VITAMIN/inorganic additive
For every kind of particle product PA that contains phytase, PB, PC, PD and PE, described goods thoroughly are mixed in the above-mentioned feed preparation of part.If particle PA, PB and PC, used mixture ratio is that to make the scope of phytase activity in the final feed preparation be the ratio of about 1500-2500 FYT/kg, and if particle PD and PE, mixture ratio is to make that the phytase activity scope is about 3000-4000FYT/kg in the final feed preparation.Use Novo Nordisk analytical method DE-9513614 as described below to measure in the feed preparation that is rich in phytase of 5 kinds of gained the sample phytase activity of each.Measure the endogenous phytase activity of the sample of the above-mentioned feed preparation that adds the goods that contain phytase in an identical manner, from the value of the various feed preparations that are rich in phytase, deduct this value.Analytical procedure: 1. analysis principle and unit definition: in Novo Nordisk analytical method ED9513614, in the aqueous solution, extract phytase from feed, then in standard conditions (37 ℃ of temperature, the acetate buffer of pH5.5, reaction times is 60 minutes, and initial phytic acid concentration is 5mM) under make the reaction of enzyme and inositol 6 phosphoric acid (phytic acid).Termination reaction is measured inorganic phosphate through the molybdate/vanadate that is added in the nitric acid.Before adding phytic acid, measure blank value through adding molybdate/vanadate/nitric acid.With the phosphoric acid salt standard substance with touchstoneization.The phytase per minute of 1FTU produces the PO of 1 μ mol under these standard conditions
4 3-2. scope: this mensuration is suitable for measuring and contains>the animal-feed phytase activity of 50FTU/kg.3. equipment list:
I) have the pH meter that the numerical value of 2 decimals shows, be equipped with suitable electrode.PH meter is used to check and regulate the pH of damping fluid and substrate solution.
Ii) be provided at the spectrophotometer that the digital reading of 3 decimals of light absorption ratio under the 415nm (optical density(OD) OD) shows, be equipped with the 10mm cuvette.
Iii) can keep 37 ± 0.2 ℃ water bath with thermostatic control.
The whizzer (the approximately typical desk centrifuge of 3000rpm) of about 2000rcf iv) can be provided.Whizzer is used for: a) be beneficial to sampling from feed extract separatin non-soluble material block, b) produce limpid solution and be used for the icrophotogrammetry measurement.
V) glass centrifuge tube.
Vi) magnetic stirring apparatus.
Vii) balance, limited amount ± 1% of weighing.
Viii) turbine mixer.
Ix): volumetric apparatus: need a certain amount of repetition suction pipe, divider and volumetric glass ware ware.Used device should transmit the amount in defined volume ± 1%.4. pharmaceutical chemicals table:
Glacial acetic acid (100%), p.a, Merck 63.
Ammonia solution 25%, p.a, Merck 5432.
Four water Ammonium Heptamolybdates, p.a, Merck 1182.
Single ammonium vanadate, p.a, Merck 1226.
Calcium dichloride dihydrate, p.a., Merck 2382.
Nitric acid 65%, p.a.Merck 456.
Potassium primary phosphate, p.a., Merck 4873
Sodium acetate trihydrate, p.a.Merck.6267.
Sodium phytate, from paddy rice, Sigma.P-3168.5. the nitric acid of reagent table 5.1. dilution: the nitric acid of 1 volume (65%) dilutes with the water of 2 volumes.Period of storage at room temperature: unlimited.5.2. Ammonium Heptamolybdate reagent: take by weighing 100.0g (NH
4)
6MO
7O
244H
2O.Be dissolved in about 800ml water.The ammonia solution that adds 10ml 25%.Adding water to final volume is 1000ml.Room temperature storage in the dark.Maximum period of storage: 8 weeks.5.3. ammonium vanadate reagent: take by weighing 2.35g NH
4VO
3Be dissolved in fully in the 400ml water that is preheating to 50-60 ℃.The nitric acid (seeing top 5.1) that adds the 20ml dilution then.Adding entry to final volume is 1000ml.Store under the room temperature in the dark.Maximum period of storage: 8 weeks.5.4.MoV termination reagent: the ammonium vanadate reagent (seeing top 5.3) that adds 1 volume in the ammonium molybdate reagent of 1 volume (seeing top 5.2).The nitric acid (seeing top 5.1) that adds 2 volume dilution then.Mixture is at room temperature stored.Every day prepared fresh.5.5. acetate buffer, pH5.5: approximately dissolve 150.1g sodium acetate trihydrate and 0.735g CaCl in the 4500ml water
22H
2O.Transfer pH to 5.50 ± 0.02 with acetate (approximately 10ml glacial acetic acid).Adding water to final volume is 5000ml.Store under the room temperature.Maximum 1 week of period of storage.Use and checked and transferred if desired pH the same day.5.6.10% calcium chloride solution: take by weighing 100g CaCl
22H
2O is soluble in water.Add water to final volume 1000ml.Be stored in room temperature.The longest period of storage: unlimited.5.7. phytic acid substrate solution: take by weighing the 1.40g sodium phytate.Be dissolved in the 200ml acetate buffer (seeing top 5.5).At room temperature transfer pH to 5.50 ± 0.02 through the acetate (corresponding to about 0.4ml glacial acetic acid) that adds dilution.Be diluted to final volume 250ml with acetate buffer (seeing top 5.5).Suppose that sodium phytate is a decahydrate, molecular weight is 1104, and this substrate solution contains the 5.1mM phytic acid so.Prepare fresh solution every day.5.8. phosphoric acid salt is stored standardized solution (50mM), approximately 10g KH
2PO
4Drying is 2 hours in 105 ℃ vacuum oven.Be stored in the moisture eliminator.Accurately take by weighing 682mg exsiccant KH
2PO
4Be dissolved in the 100ml acetate buffer (seeing top 5.5).Be stored in 0-5 ℃.The longest period of storage: 1 week.5.9.10mM phosphoric acid standard: 10ml phosphoric acid salt storage liquid standardized solution (5.8) is diluted to 50ml with acetate buffer (seeing top 5.5).Prepare fresh solution every day.6. feed sample: for example use the family expenses coffee mill that the representative sample of feed is clayed into power.7. method:
7.1. feed extracts: take by weighing 40.0 ± 0.25g feed powder.Adding 500ml contains the deionized water of 16ml 10% calcium chloride solution (seeing top 5.6).Vigorous stirring 60 minutes.Shifting about 5ml advances in the glass centrifuge tube.Centrifugal with 3000rpm, for example, 5 minutes.Prepare a series of glass test tubees.Each feed sample needs 4 test tubes (note these test tubes will be centrifugal after the enzyme reaction stage-therefore should select its size and quality).Sucking-off 100 μ l centrifugal feed samples enter in 4 by all means every pipes.
7.2. analyze: begin routine analyzer when after beginning sample extraction program, being no more than 90 minutes.2 test tubes are used for enzyme reaction, and 2 as blank.Also prepared 4 test tubes, respectively contained 100 μ l10mM phosphoric acid salt standard substance (seeing top 5.9), and 4 test tubes contain 100 μ l acetate buffers (seeing top 5.5).These are all handled by the mode identical with the enzyme sample tube.
The enzyme sample, phosphoric acid salt standard and damping fluid: 15 seconds at interval.At 0 time (t=0), add 3.0ml phytic acid substrate (5.7), mix and put into 37 ℃ of water-baths.In the time of t=60 minute, add 2.0ml and stop reagent (seeing above 5.4), mix and make at room temperature and leave standstill.The enzyme blank: along with all these samples begin, preparation is blank as follows: add 2.0ml and stop reagent (seeing above 5.4), then add 3.0ml phytic acid substrate (seeing top 5.7) immediately; Mix and make at room temperature and leave standstill.In the time of t=70-90 minute, with centrifugal all test tubes of 3000rpm 10 minutes.In the time of 120 minutes, make the OD that uses water as under the reference measure 415nm in the t=maximum value.8. calculate:, calculate Δ OD with duplicate calculating mean value and through deducting blank value for every kind of sample and phosphoric acid salt standard substance
415(blank value of phosphoric acid salt standard substance is measured values of buffer sample).
The concentration of 10mM phosphoric acid salt standardized solution: " P " mM is (from used KH
2PO
4Weight and extent of dilution).
PO in the phosphoric acid salt standard test
4 3-Mmol: " P * 0.1 ".
Δ OD
415Phosphoric acid salt standard: " A "
Sample in mensuration heavy (from weighing the extent of dilution of use and final volume): " E " g.
The Δ OD of sample
415: " B ".
Phosphoric acid salt μ mol calculation sample phytase activity with every g sample per minute release.The sample phytase activity, FTU/g=(B * P * 0.1)/(A * E * 60).8.1. example: add 16ml CaCl at 500ml water
2Extract the 40.2g feed in the solution.Feed concentration=0.0779g/ml or 0.00779g/100 μ l.
OD
415Feed specimen test=0.506
OD
415Feed sample blank=0.252
Δ OD
415Feed sample=0.254.
Actual concentrations=the 9.62mM of phosphoric acid reference liquid
Phosphoric acid salt among the 100ml=0.962 μ mol
OD
415Feed specimen test=0.563
OD
415Feed sample blank=0.197
Δ OD
415Feed sample=0.366.
Feed sample phytase activity=(0.254 * 0.962)/(0.366 * 0.00779 * 60)=1.428FTU/g.9. note: 9.1. vanadate toxicity: vanadate (vanadium of+5 valency states of oxidation) has severe toxicity (the oral LD in rat
50Be 18mg/kg).MoV stops reagent and contains the 0.6mg/ml vanadate.Final testing liquid (need great majority to handle, comprise centrifugal) contains the 0.24mg/ml vanadate (with about 2%HNO
3).9.2. in feed is measured, add calcium: comprise calcium at the aforesaid method that is used for the feed sample and add.The adding of calcium has reduced the production of free phosphoric acid during sample preparation steps,, has reduced blank and enzyme OD value that is.9.3. phosphoric acid salt standard substance: up at least 1.5 OD
415The time phosphate reaction be line style.Phosphoric acid salt standard substance observed value repeatability is fabulous, only uses 1 phosphate concn to be enough to produce accurate calibration.The Δ OD of 10mM phosphoric acid salt standard substance
415Near 0.37.9.4. maximum sample concentration: OD
415Should be no more than 0.8.If obtain higher value, can advise regulating the diluted sample degree.For example, use 1000ml deionized water replaces the 500ml (seeing above) in 7.1.Steam heating:
The feed a part preparation that respectively is rich in the feed preparation of phytase and does not add phytase for part carries out the steam heating program, and is wherein used similar in the condition of steam heating program and the typical feed pelletizing process: transfer to described 100g feed preparation in the B (Schott Duran 21,341 44 05) and transmit steam by funnel and also heat by the feed preparation that holds in funnel thus.Steam heating continues 30 seconds, evenly stirs feed therebetween.In all strokes the supply of steam keep constant and be enough to make the temperature of feed preparation when heating finished in 30 seconds reach>85 ℃.Then the feed preparation of heating is poured out funnel and is cooled to envrionment temperature.
Use Novo Nordisk analytical method ED-9513614 (seeing above) measures the phytase activity of the sample of each steam-heated feed preparation, and proofreaies and correct each steam-heated activity that is rich in the goods of phytase through deducting the remaining endogenous phytase activity that the steam-heated feed preparation that does not add the phytase goods is measured.
The percentage ratio that phytase activity keeps in each steam-heated feed preparation that is rich in phytase provides in following table: the reservation (%) that adds the phytase activity in the feed preparation contains the particle PA 14PB 14PC 68PD 56PE 39 of phytase
These results show that (both all are article according to the invention and have high-load water-fast material for use particle PC or particle PD, comprise hydrophobic substance) cause the reservation of enzymic activity in the steam heating process to be significantly higher than to particle PA, the viewed active reservation of PB and PE (these compositions are outside scope of the present invention).
Embodiment 4
Steam heating is to the influence of the feed preparation xylanase activity that contains zytase: check to mix respectively to contain zytase particle product XA, XB, XC, the reserving degree of enzymic activity after the feed preparation steam heating of one of XD and XE (seeing embodiment 2).Cavings feed preparation with (seeing above) the identical essentially consist described in the embodiment 3 is used for this purpose.
For every kind of particle product that contains zytase, it is about 800-1000FXU/kg that described goods are mixed in the feed preparation to produce final xylanase activity scope.If particle XA and XB, be mixed in the 10kg feed preparation through the grain amount that will be equivalent to 96000FXU and carry out; The feed that then the part gained is rich in enzyme thoroughly mixes with 110kg basal feed preparation, and producing altogether, the calculated activity of 120kg is the material of 800FXU/kg.If particle XC, XD and XE, preparation is than short run (<1kg) enrichment feed preparation.
Use Novo Nordisk analytical method ED-9511460.2 described below to measure in the feed preparation that is rich in xylan of 5 kinds of gained the sample xylanase activity of each.Analytical procedure:
1. analysis principle: in Novo Nordisk analytical method ED-9511460.2, enter the aqueous solution from feed sample extraction zytase.Make the wheat xylan reaction of this enzyme and modification then.This wheat xylan be crosslinked (make its soluble) and through and blue dyes coupling dyeing, and it can obtain with tablet form.Under normalization condition (temperature is 70 ℃, pH6.0, the reaction times is 120 minutes), react.Stop through increasing the pH value then.Filtering solution and with the blueness of metric measurement solution, it is enzymatic substrate dissolved observed value.Measure through not enzyme-added sample and to measure blank value.Through add this test of the active calibration of known enzyme to the feed sample.Make the increase of reaction be used to calculate the activity of feed sample owing to adding enzymic activity.
2. scope: this test is suitable for measuring the feed Bio-Feed that contains 50-250FXU/kg
TMWheat L xylanase activity.If regulate concentration and extent of dilution, also can measure higher activity.This paper comprises the indication of every kg field of activity 200-1000FXU.
3. equipment list: i) pH meter, the digital reading with 2 decimals shows, is equipped with suitable electrode.PH meter is used to check and regulate the pH of buffered soln.Ii) spectrophotometer is provided at the digital reading demonstration of 3 decimals of absorbancy under the 585nm (optical density(OD) OD), is equipped with the 10mm cuvette.Iii) the laboratory desk centrifuge is used for separating the insoluble substance piece of feed extract to help sampling.Iv) be used to contain the v) water bath with thermostatic control of centrifuge tube of 5-10ml sample, can maintain 70 ± 0.5 ℃.Vi) vortex mixer.Vii) magnetic stirring apparatus.Viii) glass fibre filter, Whatman GF/C, 9cm diameter.Ix) balance can be weighed limited amount in ± 1%.X) displacement apparatus: need a certain amount of repetition suction pipe, divider and volumetric glass ware ware.Used device should transmit the amount of defined volume ± 1%.Xi) glass test tube, for example 15mm * 100mm has lid.4. pharmaceutical chemicals table:
The phosphate dihydrate disodium hydrogen, p.a, for example Merck 6580.
Sodium dihydrogen phosphate-water, p.a, for example, Merck 6346.
Three (methylol) methane, for example, Sigma 7-9.Sigma T-1378.
Zytase AX, zytase measure tablet, Australian Megazyme Pty company limited.
Bio-Feed
TMWheat L (the zytase goods of tool known activity), Novo NordiskA/S, Denmark.
5. reagent table: 5.1. phosphate buffered saline buffer, pH6: with 60.0g NaH
2PO
4H
2O and 11.6gNa
2NPO
42H
2O is dissolved in about 4500ml water.Transfer pH to 6.00 ± 0.05 with 1N HCl or 1N NaOH.Being diluted to final volume is 5000ml.Store under the room temperature.Maximum period of storage: 1 week.5.2.Tris termination reagent: 10.0g three (methylol)-methane is dissolved in the 1000ml water.Store under the room temperature.Maximum period of storage: 1 week.5.3. enzyme standardized solution: from Bio-Feed
TMThe buffered soln in the 4.8-5.2FXU/ml scope that wheat L (having known xylanase activity) preparation has accurate known activity.For example, if Bio-Feed
TMWheat L product has 500FXU/g, and about 1000mg and being dissolved in the 100ml phosphate buffered saline buffer (seeing top 5.1) accurately weighs.At room temperature store.The longest period of storage: 2 hours.6. feed sample: " feed sample " is the intact sample of the feed received of laboratory.
Be used for Bio-Feed
TMThe sample F X:FX that the wheat zytase is measured is that 100g takes from the sample of " feed sample ".The program that is used to take out FX must guarantee that it is representational sample.FX is clayed into power, be used for this mensuration then.Replication for to identical " feed sample " should repeat the gross sample program.
7. method: take by weighing 2 samples of feed FX, each 40 ± 0.4g.Be labeled as " a " and " b ".In program subsequently, handle two samples in an identical manner.In the 0 o'clock time (t=0); Add 800ml water and begin and stir.At time t=30 minute, get about 5ml and also shift in the centrifuge tube.Be labeled as " aS " and " bS ".
The enzyme standardized solution that adds 1.0ml (sees 5.3, above) and continuously stirring.At time t=45 minute, get about 5ml and also shift in the centrifuge tube.Be labeled as " aE " and " bE ".With about 3000rpm centrifugal all 45 minutes by all means.
Prepare 16 glass test tubees, respectively contain 2.0ml phosphate buffered saline buffer (seeing above 5.1).From 1 to 16 continued labelling.These test tubes use as follows:
Test tube 1-3: working sample " aS "
Test tube 4: blank
Test tube 5-7: working sample " aE "
Test tube 8: blank
Test tube 9-11: working sample " bS "
Test tube 12: blank
Test tube 13-15: working sample " bE "
Test tube 16: blank
200 μ l feed extract centrifugates are shifted in separately the mensuration test tube.Mix with vortex mixer.In blank pipe, add 200 μ l phosphate buffered saline buffers (seeing above 5.1).
Sample extraction begins after 60-90 minute the following mensuration program of beginning: use the timed interval easily between each pipe, add 1 zytase AX16 in every pipe by all means, build test tube and it is positioned in 70 ℃ of water-baths.It is important mixing never in any form behind the attention adding tablet.
In water-bath after 120 minutes, test tube is handled as follows: take out test tube, add 5ml Tris and stop reagent (seeing top 5.2), build and with vortex mixer thorough mixing content again.All test tube room temperatures were left standstill 15 minutes, and then mix and filter in the into clean test tube by Whatman GF/C filter.Measure the OD value of all 16 samples under 585nm.Filter in back 30 minutes and finish these measurements.8. calculate:
Calculate " aS " " bS ", the OD that " aE " and " bE " measures
585Mean value (respectively being the average of 3 observed values).Calculate blank OD
585Mean value.Deduct average blank value through the corresponding average measurement value of associating and calculate " aS ", " bS ", the Δ OD of " aE " and " bE ".
Calculate (with the FXU/kg feed) adds the feed sample in leaching process enzymic activity (for example, have in the 1ml adding 40g feed of 5FXU/ml produce 125FXU/kg).
Calculate the activity of feed sample " a " and " b "
The FXU/kg feed, sample " a "=
(Δ OD, aS) * (added enzyme, FXU/kg)/[(Δ OD, aE)-(Δ OD, aS)]
The FXU/kg feed, sample " b "=
(Δ OD, bS) * (enzyme of adding, FXU/kg)/[(Δ OD, bE)-(Δ OD, bS)]
The xylanase activity of " feed sample " provides with the mean value of " a " and " b ".9. the indication that has the feed sample of 200-1000FXU/kg
The concentration of enzyme standardized solution (seeing top 5.3) is increased to 20FXU/ml from 5.The volume of feed extract centrifugate used in measuring is reduced to 50 μ l from 200 μ l.Need not to carry out other modification.Pill/steam heating
If use particle XA and XB preparation to contain the feed preparation of zytase respectively, use guiding scale steam regulator (being equipped with the continuous horizontal mixing tank of lodicule and steam-in) with this preparation pill.Flow velocity by setter is 200kg/ hour, and the residence time in setter is 30 seconds, and it is 95 ℃ that the steam regulation input makes the vapor temperature of output feed.Feed enters Simon Heesen squeezer and extruding from the outlet of setter makes its mould by having 3mm * 35mm hole size to become piller.The piller sample transfer is cooled to envrionment temperature in water cooler and in air.The sample of getting the cooling piller is used for the xylanase activity analysis.
If use particle XC respectively, the feed preparation that XD and XE preparation contain zytase adopts and steam heating program similar described in the embodiment 3.The supply of steam maintenance is constant and last between 30 second heating period in all strokes is enough to that the temperature of feed preparation is reached>and 90 ℃.Then the feed preparation of heating is poured in the funnel and is cooled to envrionment temperature.
Use Novo Nordisk analytical method ED 9511460.2 (seeing above) to measure the sample xylanase activity of each steam-heated feed preparation.
The percentage that xylanase activity keeps in the feed preparation that is rich in zytase that each Steam Heating is crossed provides in following table: the xylanase activity (%) that adds the reservation in the feed preparation contains the particle XA 35XB 93XC 10XD 40XE 85 of zytase
These results show: (they all are goods of the present invention and have high-load water-fast material to use particle XB and XE, comprise hydrophobic substance) cause in pelletizing process during the steam heating or in other steam heating process the reservation of enzymic activity be higher than at particle XA, the viewed enzymic activity of XC and XD (said composition is outside scope of the present invention) keeps.
Claims (16)
1. a particulate state contains enzyme preparation, comprises: one or more hydrophobic substances that are total at least 1% weight (w/w); Be total to one or more water-fast materials of 75%w/w at least, comprise said hydrophobic substance.
2. the goods of claim 1 comprise the said hydrophobic substance of 5%w/w at least.
3. according to the goods of claim 1 or 2, comprise the said water-fast material of 90%w/w at least.
4. according to each goods among the claim 1-3, comprise the said hydrophobic substance that is uniformly distributed in the 5-95%w/w in the said product grain material basically.
5. according to each goods among the claim 1-3, comprise and contain the enzyme core and contain said hydrophobic substance and surround the said coatings that contains the enzyme core.
6. according to the goods of claim 5, comprise the said hydrophobic substance of 1-50%w/w.
7. according to the goods of claim 5 or 6, comprise the said hydrophobic substance of 5-15%w/w.
8. according to each goods of aforementioned claim, comprise be selected from inorganic salt and polysaccharide material as water-fast material.
9. according to each goods among the claim 5-7, comprise be selected from inorganic salt and polysaccharide material as water-fast material.
10. according to the goods of claim 9, wherein saidly contain the Mierocrystalline cellulose that the enzyme core comprises fibers form.
11. according to each goods among the claim 1-10, wherein said enzyme is to be selected from phytase, phosphoesterase, zytase, beta-glucanase, alpha-galactosidase, α-Dian Fenmei, beta-amylase, cellulase, a kind of enzyme of polygalacturonase and peptase.
12. produce the method that contains the enzyme animal feedstuff compositions, wherein each the mixture that contains other composition in enzyme preparation and the final feed composition among the claim 1-11 carried out the steam heating step.
13., comprise step with said final feed composition pill according to the method for claim 12.
Contain application in the enzyme animal feedstuff compositions 14. will contain enzyme preparation in production according to each particulate state among the claim 1-11.
15. goods that use among the claim 1-11 each contain the enzyme animal feedstuff compositions as what contain the production of enzyme initial substance.
16. can contain the enzyme animal feedstuff compositions by what the method for claim 12 or 13 obtained.
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JP (1) | JPH11514240A (en) |
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WO (1) | WO1997016076A1 (en) |
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Publication number | Priority date | Publication date | Assignee | Title |
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DK13491D0 (en) * | 1991-01-25 | 1991-01-25 | Novo Nordisk As | APPLICATION OF AN ENZYMOUS GRANULATE AND PROCEDURE FOR PREPARING A TABLET FORM |
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1996
- 1996-11-01 AU AU74898/96A patent/AU7489896A/en not_active Abandoned
- 1996-11-01 WO PCT/DK1996/000455 patent/WO1997016076A1/en not_active Application Discontinuation
- 1996-11-01 CN CN96197980A patent/CN1201372A/en active Pending
- 1996-11-01 JP JP9516999A patent/JPH11514240A/en active Pending
- 1996-11-01 EP EP96937191A patent/EP0858266A1/en not_active Withdrawn
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN105211639A (en) * | 2004-09-27 | 2016-01-06 | 诺维信公司 | Enzyme granulate |
CN101668518B (en) * | 2007-02-14 | 2014-05-07 | 尼尔·A·所罗门 | A lactase formulation |
CN101120730B (en) * | 2007-09-13 | 2010-07-28 | 武汉工业学院 | Paddy special-purpose compound enzyme feed additive |
CN104204179A (en) * | 2011-06-20 | 2014-12-10 | 诺维信公司 | Particulate composition |
CN107475235A (en) * | 2011-06-20 | 2017-12-15 | 诺维信公司 | Particulate composition |
Also Published As
Publication number | Publication date |
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WO1997016076A1 (en) | 1997-05-09 |
AU7489896A (en) | 1997-05-22 |
JPH11514240A (en) | 1999-12-07 |
EP0858266A1 (en) | 1998-08-19 |
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