CN1671294A - Feed for fry young fishes and method of producing hydrolyzate of low-phytin vegetable protein to be used therein - Google Patents

Feed for fry young fishes and method of producing hydrolyzate of low-phytin vegetable protein to be used therein Download PDF

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Publication number
CN1671294A
CN1671294A CNA038177072A CN03817707A CN1671294A CN 1671294 A CN1671294 A CN 1671294A CN A038177072 A CNA038177072 A CN A038177072A CN 03817707 A CN03817707 A CN 03817707A CN 1671294 A CN1671294 A CN 1671294A
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feed
phytic acid
low
protein
hydrolyzate
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CN100379354C (en
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中森俊宏
古田均
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Fuji Oil Co Ltd
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Fuji Oil Co Ltd
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    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K50/00Feeding-stuffs specially adapted for particular animals
    • A23K50/80Feeding-stuffs specially adapted for particular animals for aquatic animals, e.g. fish, crustaceans or molluscs
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/14Vegetable proteins
    • A23J3/16Vegetable proteins from soybean
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23JPROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00Working-up of proteins for foodstuffs
    • A23J3/30Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/346Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K10/00Animal feeding-stuffs
    • A23K10/10Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/14Pretreatment of feeding-stuffs with enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/142Amino acids; Derivatives thereof
    • A23K20/147Polymeric derivatives, e.g. peptides or proteins
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/153Nucleic acids; Hydrolysis products or derivatives thereof
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K20/00Accessory food factors for animal feeding-stuffs
    • A23K20/10Organic substances
    • A23K20/189Enzymes
    • AHUMAN NECESSITIES
    • A23FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23KFODDER
    • A23K40/00Shaping or working-up of animal feeding-stuffs
    • A23K40/30Shaping or working-up of animal feeding-stuffs by encapsulating; by coating
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02ATECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/80Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
    • Y02A40/81Aquaculture, e.g. of fish
    • Y02A40/818Alternative feeds for fish, e.g. in aquacultures

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Food Science & Technology (AREA)
  • Zoology (AREA)
  • Animal Husbandry (AREA)
  • Biochemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Physiology (AREA)
  • Biomedical Technology (AREA)
  • Biotechnology (AREA)
  • Insects & Arthropods (AREA)
  • Birds (AREA)
  • Marine Sciences & Fisheries (AREA)
  • Nutrition Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Fodder In General (AREA)
  • Feed For Specific Animals (AREA)

Abstract

It is intended to provide a feed which is appropriate for enhancing the survival rate of young fry fishes with underdeveloped digestion and absorption functions or promoting the growth of them. It is also intended to obtain a vegetable protein hydrolyzate to be used in this feed. To obtain a low-phytin vegetable protein hydrolyzate, a process for producing the low-phytin vegetable protein hydrolyzate which comprises (a) the step of digesting a protein with the use of a protease and (b) the step of digesting phytic acid with the use of an enzyme digesting phytic acid has been completed. In the case where the vegetable protein is soybean protein, for example, it is preferable that the low-phytin soybean protein hydrolyzate has an average molecular weight of from 200 to 10,000. It is also preferable that the phytic acid content is 0.05% by weight or less (based on dry solid matters). A feed for young fry fishes containing this low-phytin soybean protein hydrolyzate makes it possible to provide a feed for young fry fishes which promotes the growth of young fry fishes and enhances the survival rate.

Description

Feed for fry young fishes and be used for the manufacture method of hydrolyzate of low-phytin vegetable protein wherein
Technical field
The invention provides the feed for fry young fishes that uses the hydrolyzate of low-phytin vegetable protein that has reduced phytic acid.The invention still further relates to the manufacture method that this has reduced the hydrolyzate of low-phytin vegetable protein of phytic acid.
Background technology
In the past, used soybean material (soya-bean cake, soya-bean milk, soybean protein etc.) as the protein raw materials of breeding fish with feed always.
In addition, if knownly remove phytic acid contained in the soybean, can improve the efficient of feed.
As the invention of removing or decomposing phytic acid, the feed material (spy opens flat 9-140334 communique) of soybean origin such as known (a) feed (spy opens flat 11-000164 communique, the spy opens flat 8-205785 communique), (b) defatted soybean, soybean residue, (c) soybean protein (the application applicant's spy opens the 2000-300185 communique, the spy opens communique No. 4503002) and (d) soya-bean milk (spy that the spy opens according to 59-166049 communique, the application applicant opens the 2000-245340 communique) etc.
But the low-phytin hydrolysate is also of no use is breeding fish with in the feed.
On the one hand, the applicant is studying always and is using the protolysate that obtains with the soy proteins raw material to use feed as breeding fish.For example, spy invention, the spy of fish meal that open the survival rate of the raising juvenile fish seedling of putting down in writing in the flat 7-227223 communique opens invention of putting down in writing in the flat 8-51937 communique etc.In addition, in order further to improve the survival rate of juvenile fish seedling, the improvement of soybean protein hydrolyate is also studied.
On the other hand, about soybean protein hydrolyate, the application applicant discloses and has been used for hydrolyzate of low-phytin vegetable protein (spy the opens flat 8-092123 communique) nephrotic, remove phytic acid with resin.
Again on the one hand, known inoculation aspergillus makes the soybean protein fermentation, carries out protein breakdown and phytase treatment, makes low-phytin protolysate (spy opens flat 9-023822 communique).
Others, following invention is arranged, when being enzyme processing soybean protein raw material, owing to use thick enzyme, protease plays a part identical with phytase, perhaps protease is decomposed and the invention of phytase treatment combination, but do not disclose, also do not instruct zymolytic soybean protein medium content (spy opens clear 51-125300 communique, the spy opens 2000-51706 communique etc.) that is used to breed fish.
As mentioned above, the applicant uses soybean protein hydrolyate in continuous research in breeding fish with feed, is also studying the low-phytin soybean protein hydrolyate always, but and does not know they are used in and breed fish with feed particularly in the juvenile fish seedling.
In addition, the method of removing the phytic acid of soybean comprises that (1) remove method (spy opens flat 8-173052 communique, the spy opens flat 9-121780 communique etc.), (2) of phytic acid and separate the method for phytic acid, the method (spy opens the 2001-163800 communique) that remove with absorption such as resins (3) etc. with the phytase five equilibrium in the water that uses salt is put forward the process of soybean protein, compare with the method for (3) with (1), the operation of method (2) is uncomplicated, industrial be favourable.
In addition, separate the object of the method for phytic acid with the phytase five equilibrium about these (2), what (a) know is that soybean, defatted soybean as purposes such as feeds etc., (b) also known soya-bean milk, soybean protein isolate etc. are several, but not too knows about (c) vegetable protein hydrolyzate mixture.
The feed material (spy opens flat 9-140334 communique) of soybean origin such as known for example (a) feed (spy opens flat 11-000164 communique, the spy opens flat 8-205785 communique), (b) defatted soybean, soybean residue, soybean protein (the application applicant's spy opens the 2000-300185 communique, the spy opens communique No. 4503002) and soya-bean milk (spy that the spy opens clear 59-166049 communique, the application applicant opens the 2000-245340 communique) etc.
But, but not known about (c) low-phytate calcium enzyme vegetable protein hydrolyzate, open the application applicant's spy and to disclose the method for using resin in the flat 8-092123 communique.
In addition, open in the flat 9-023822 communique, disclose the inoculation aspergillus and made the soybean protein isolate fermentation, carry out enzyme simultaneously and decompose and phytase treatment, obtain the method for the low peptide product of phytic acid content the spy.
But, also do not make phytic acid as described in the present invention in the method that detects below the boundary.
Summary of the invention
The inventor etc. find in studying, the feed of using for the fish that survives after using vegetable protein hydrolyzate as the less developed small fish of digestion and absorption function, fry, birth, the peptide product is sufficient inadequately, need the property digested and assimilated excellence and the few vegetable protein hydrolyzate of phytic acid content.Therefore, the objective of the invention is to obtain the few low-phytate vegetable protein hydrolyzate of phytic acid content, especially preferably obtain phytic acid at the low-phytate vegetable protein hydrolyzate that detects below the boundary.
Further purpose is to use the feed for fry of this low-phytate vegetable protein hydrolyzate.
The inventor etc. found that the contained phytic acid of soybean protein hydrolyate that uses by reducing by constantly concentrated research in feed, can improve the survival rate of juvenile fish seedling, can promote growing of fry, thereby finish the present invention.
In addition, by following discovery, obtain hydrolyzate of low-phytin vegetable protein.
That is to say that the applicant has finished the low-phytin protolysate (spy opens flat 8-092123 communique) that obtains by resin treatment earlier, but for phytic acid content is being detected below the boundary, resin treatment is pretty troublesome.
On the other hand, the applicant uses the phytase treatment soybean protein, make low-phytate soybean protein (spy opens the 2000-300185 communique), but this low-phytate soybean protein of enzymolysis only, it is very low also to be difficult to obtain phytic acid content, promptly at the soybean protein hydrolyate that detects below the boundary.
Therefore, further concentrated research is found at first hydrolysate of soybean protein to carry out phytase treatment afterwards to specific molecular weight ranges, can obtain the soybean protein hydrolyate of phytic acid considerably less (it is following to detect boundary).
Also find when enzymatic hydrolysis of soybean albumen, after soybean material water extraction soybean protein, just do not carry out enzymolysis, carry out phytase treatment earlier, carry out enzymolysis then, also can remove phytic acid to detecting below the boundary if do not carry out drying.
Make us is that these phytic acid contents are that the extremely few like this soybean protein hydrolyate of phytic acid content is compared the local flavor excellence with a soybean protein hydrolyate of process enzymolysis detecting below the boundary more unexpectedly.
The present invention is based on that these discoveries finish.
That is to say that feed for fry young fishes of the present invention is characterized in that containing the hydrolyzate of low-phytin vegetable protein that phytic acid content is 0.05 weight % or following (in dry solid content) in feedstuff.
Mean molecule quantity is that 200~10000 vegetable protein hydrolyzate is proper hydrolyzate of low-phytin vegetable protein.
In addition, the invention still further relates to the manufacture method of hydrolyzate of low-phytin vegetable protein, it is characterized in that the operation that comprises that (a) decomposes phytic acid with the operation of protease decomposing protein and the enzyme of (b) using the decomposition phytic acid.
Behind the protease decomposition of protein, preferably decompose phytic acid with the enzyme that decomposes phytic acid.
After preferably decomposing undried albumen, use the protease decomposing protein with the enzyme that decomposes phytic acid.
The enzyme preferably myo-inositol six-phosphatase that decomposes phytic acid.
The pH of phytase treatment is preferably 6~9.
The mean molecule quantity of low-phytin protolysate is preferably 200~10000.
Phytic acid content in the hydrolyzate of low-phytin vegetable protein can not detect phytic acid preferably in dry solid content with vanadium molybdic acid absorption photometry (detecting boundary 5mg/100g).
The best mode that carries out an invention
At first describe with regard to feed for fry young fishes.
The hydrolyzate of low-phytin vegetable protein that is used for feed for fry young fishes of the present invention is 0.05 weight % or following in the phytic acid content of dry solid content, preferred 0.01 weight % or following, more preferably 0.004 weight % or following (it is following to detect boundary) are proper.
The amount of contained phytic acid is few more in the vegetable protein hydrolyzate, and the survival rate of juvenile fish seedling is high more, is preferred.In addition, have the growth that promotes fry, give ight soil viscosity, anti-sealing because ight soil becomes muddy effect.
The mean molecule quantity that is used for the hydrolyzate of low-phytin vegetable protein of feed for fry young fishes of the present invention is that 200~10000 (preferred 300~5000) are proper.The low-phytin protolysate that molecular weight is big improves the effect of growing of promotion juvenile fish seedling and the weak effect of survival rate, if decrease in molecular weight is until becoming amino acid whose words, the osmotic pressure of feed raises, and becomes to be easy to dissolving, and is improper as feed.
Use feed so long as breed fish, the soybean protein hydrolyate that molecular weight ratio is bigger is also passable, but the fish of supporting is the juvenile fish seedling, little relatively good of mean molecule quantity, and especially just from the small fish of ovum hatching, the oligo peptide that preferred molecular weight is little.
Feed of the present invention as its composition, contains protein ingredient 40~70 weight %, and preferred about 50~60 weight % are proper.
Feed of the present invention contains hydrolyzate of low-phytin vegetable protein 1 weight %~30 weight %, and preferred 3~25 weight % are proper.Usually, replace with above-mentioned low-phytate vegetable protein hydrolyzate more than the 3 weight % of the protein ingredient in the feed of the present invention, preferred 10~80 weight % are proper.Substitute proportion increases, and it is difficult that the granulating of this feed becomes.
Hydrolyzate of low-phytin vegetable protein is few in the feed of the present invention, does not improve the effect of the rate of growth of the survival rate of juvenile fish seedling and fry, and is too much, can cause growth disorder, is not preferred therefore.This is that the fish of breed seedling production difficulties such as Red Snapper, flatfish has jointly, and is different with the breed of other fish.
In addition, as the protein component beyond the hydrolyzate of low-phytin vegetable protein, can merge and use krill, fish converted products, egg processed goods, milk product, gelatin, fish meal, fish Jie class extract, yeast extract, fish-egg extract.
Feed of the present invention can also contain phosphatide such as carbohydrate, fat, vitamin, mineral matter, n-3 highly unsaturated fatty acid and soybean lecithin except that containing above-mentioned hydrolyzate of low-phytin vegetable protein, other protein.
The n-3 highly unsaturated fatty acid is the necessary aliphatic acid of seawater fish such as flatfish, and feed phosphatide such as soybean lecithin are the necessary compositions of breed of juvenile fish seedling, therefore can be contained in the feed of the present invention.
Particle diameter, floatability, sinking speed and not stripping of nutrient in water that the form of feed of the present invention preferably keeps fish to be easy to absorb, in alimentary canal, digested and assimilated such form, when being used in particular for small fish such as flatfish and shrimp, preferably, make the particulate feed by microencapsulation etc.
The method of the hydrolyzate of low-phytin vegetable protein microencapsulation of mean molecule quantity 200~10000 can be adopted for example with the spray-dired method of the aqueous solution of hydrolysate.In addition, also can in order to regulate stripping property, floatability, the dispersiveness to seawater, also can add grease before spraying as required, after the spraying, behind the interpolation hardened fat, stirring and coating wait and prepare.
Feed of the present invention can be according to the age in days of juvenile fish seedling, in juvenile fish seedling stage, combine with biological feedstuff or separately, with 30 minutes~1 hour at interval, an amount of feed.
Below describe and be used for an as above manufacture method of the hydrolyzate of low-phytin vegetable protein of the feed of juvenile fish seedling.
(plant protein material)
The used vegetable protein of the present invention can use known vegetable protein, and the albumen of cereal, glyceride stock obtains easily, and particularly soybean protein is industrial can produce in a large number, is one of preferred protein raw materials therefore.
The following describes the example that uses the soybean protein that contains some phytic acids, but this method also is the method that can be applied in other vegetable protein.
The soybean protein raw material that is used to make hydrolyzate of low-phytin vegetable protein of the present invention can comprise that soya-bean milk (also comprises the degreasing soya-bean milk.As follows) as long as, concentrate soya-bean milk, soy protein concentrate, soybean protein isolate, defatted soybean etc. and comprise soybean protein.
Defatted soybean does not preferably have albuminous degeneration or the slight protein-denatured low sex change defatted soybean of handling through processing of what is called, is not subjected to qualifications such as kind, the place of production.In general, proper as raw material as the defatted soybean that the extraction solvent carries out low temperature extraction processing n-hexane, particularly NSI (but nitrogen solutbility coefficient) is more than 60, and preferred low sex change defatted soybean more than 80 is relatively good.
(enzymolysis of soybean protein)
The enzyme solution of soybean protein can use soy proteins in aqueous solution (soybean protein slurry or solution).
For example, in the occasion of using low denatured soybean protein, the concentration that is used for the soy bean proteinous soln of enzyme processing is 1 weight %~30 weight %, preferred 5~15 weight %, and more preferably 8~12 weight % are proper.This concentration is low not to be hindered enzymolysis yet, but the productive rate reduction is not preferred.
Be used for protease of the present invention (protease) and can separately or merge extracellular protease or the intracellular protein enzyme that use derives from animal, plant or microorganism.Can use serine protease (subtilopeptidase A of the trypsase of animal origin, chymotrypsin, microorganism origin, carboxypeptidase etc.), thiol proteinase (papain of plant origin, ficin, bromelain etc.), carboxyl protease (pepsin of animal origin etc.) particularly.Further be in particular " ア Network チ Na-ゼ " (the scientific research pharmacy strain system) of " プ ロ チ Application FN " (big and change into strain system), the grey chain enzyme bacteria origin of aspergillus oryzae origin, from " ア Le カ ラ-ゼ " (ノ ボ society system) of bacillus licheniformis, from hay bacillus " プ ロ チ Application A " (big and change into strain system).In addition, as the enzyme that contains the intracellular protein enzyme, can enumerate " the プ ロ テ ア one ゼ S ", big and change into " プ ロ チ Application AC-10 " and " the PVC オ プ ラ-ゼ (Na ガ ゼ Seikagaku Kogyo Co. Ltd.) etc.;, can enumerate " プ ロ テ ア one ゼ M " that amano pharmaceutical (strain) is made of (strain) system of amano pharmaceutical (strain) system as the protease that contains the outer and intracellular protein enzyme of born of the same parents.
What are different because of the kind of the protease that uses for hydrolysising condition of the present invention, and in general, preferably under the action pH scope of this protease, operative temperature, optimum reacting time, the enzyme that uses capacity is with hydrolytic soya bean protein.Consider the occasion of purposes of the food (for example through feeding tube nutriment etc.) of lipid-metabolism-improving agent and restriction salinity at the same time, if pH is 5~10, preferred pH6~9 can be reduced the salt that neutralization generates, thereby are preferred.
For the degree of hydrolysis, mean molecule quantity is 200~10000, and preferred 300~5000 is proper.The degree that can regulate hydrolysis according to purpose and purposes.For example, in the occasion of foodstuff and feed, bigger molecular weight is also harmless, be used as the breed fish occasion of using feed and the occasion that is used as feed for fry young fishes, preferably be easy to the low-molecular-weight that digests, suitable mean molecule quantity is 200~5000, more preferably is 200~2000.
(decomposing phytic acid) with the phytic acid catabolic enzyme
As the enzyme that is used for decomposition phytic acid of the present invention, can use enzyme from plants such as wheat and potatos, perhaps from the enzyme of plucks such as intestinal tube, from microorganisms such as bacterium, yeast, fungi, actinomyces and enzymes such as phytase that have the phytic acid degrading activity and phosphatase.
As the enzyme that decomposes phytic acid, phytase and phosphatase are proper, more preferably phytase.Phytase can use various bacterial strains with product phytase ability such as aspergillus, rhizopus, saccharomyces, mucor, Geotrichum.Preferably proper from the enzyme of aspergillus, aspergillus (Aspergillus) more preferably: optional from from the phytase of fig aspergillus (Aspergillus ficuum), from the phytase of aspergillus niger (Aspergillus niger) and the phytase of native inulinase (Aspergillus terreus).For the phytic acid in the soybean is decomposed into inositol, need to cut off ester group, the enzyme that cuts off is a phytase.
In addition, also can use acid phosphatase as the Mycophyta origin of acid phosphatase.That is to say, can be selected from the acid phosphatase of fig aspergillus (Aspergillus ficuum) origin, the acid phosphatase of aspergillus niger (Aspergillus niger) origin and the acid phosphatase of native inulinase (Aspergillus terreus) origin.
The decomposition reaction of the phytic acid that enzyme is handled can be implemented under condition as mild as a dove, and is therefore extremely little to the influence of protein.For example, enzyme reaction of the present invention is carried out getting final product in 0.1~30 hour under 30~60 ℃.
In the present invention, the pH particular importance during the phytic acid decomposition reaction, preferred 6.2~8.5 in pH6~9, further 6.2~7 implement relatively good.Be lower than its dissolubility reduction of soybean protein of handling under the pH6.0, the local flavor variation is not preferred.In addition, pH surpasses 9.0, and local flavor is variation also, neither be preferred.In above-mentioned pH scope, decompose phytic acid, can make the soybean protein that has reduced phytic acid better.
Thereby the enzyme that be fit to use the in the present invention preferably neutrality more than pH6 and even alkaline pH scope can decompose the enzyme of phytic acid and phytate, and there is no particular limitation to its source, can use above-mentioned enzyme.
Use enzyme under can no matter Powdered, the liquid form, with respect to the crude protein weight in the soybean protein, with 0.01~10 weight %, preferred 0.05~2 weight %, more preferably 0.1~1 weight % adds other and implements, but tire as enzyme, add the thick legumin of 0.1~100U/g, the thick legumin of preferred 0.5~20U/g, more preferably the phytase of the thick legumin of 1~10U/g is relatively good.In addition,,, make by 0.5ml to contain the reactant liquor reaction 30 minutes that 0.2Mtris-HCl buffer solution (pH6.5), 0.4ml distilled water and the 0.1ml enzyme liquid of 4mM sodium phytate forms, add 1.0ml 10%TCA and also stop reaction at 37 ℃ for enzymatic activity.With the Fiske-Subbarow method inorganic phosphate content in this reactant liquor is carried out quantitatively.Under these conditions, the free enzyme amount of the inorganic phosphate that in 1 minute, makes 1 μ mol as 1 unit (U).
In the present invention, as long as comprise with the operation of protease decomposing protein and the operation of decomposing phytic acid with the enzyme that decomposes phytic acid, its order combination of sample whatsoever all can, through these operations, can make low-phytinization, be in the vegetable protein hydrolyzate phytic acid content with dry solid content count 0.5% or below, preferred 0.2% or below.And, in order to make phytic acid content, behind the enzyme decomposing protein, carry out phytase treatment detecting below the boundary 5mg/100g, be more preferably for making hydrolyzate of low-phytin vegetable protein.On the other hand, protein carried out phytase treatment after, even enzymolysis protein matter also is difficult to make phytic acid content detecting below the boundary 5mg/100g.But, if state the not occasion of denatured soybean protein before use, after making aforementioned phytase act on this soy bean proteinous soln (not having powder for drying) decomposition phytic acid, carry out aforesaid protease and decompose, can obtain phytic acid content at the required hydrolyzate of low-phytin vegetable protein that detects below the boundary.
The mean molecule quantity of the hydrolyzate of low-phytin vegetable protein that as above obtains is 200~10000, and preferred 300~5000 is proper.
In addition, phytic acid content is in the dry solid content of hydrolyzate of low-phytin vegetable protein, with vanadium molybdic acid absorption photometry (detecting boundary 5mg/100g), detection be 0.5% or below, preferably do not detect phytic acid.
(embodiment)
Below by embodiment embodiments of the present invention are described.
At first, feed is described.
(Production Example 1) (manufacturing of low-phytate soybean protein hydrolyate)
In 10 weight portion defatted soybeans, add 7 times water, under 50 ℃, pH7, stir to extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, with sulfuric acid soya-bean milk is adjusted to pH4.5 after, by centrifugation etc., after being separated into isoelectric point precipitates albumen matter and whey protein, with respect to isoelectric point precipitates albumen matter, add 4 times of water after, be adjusted to pH6.0 with NaOH, prepare soy bean proteinous soln 50 weight portions of 8% concentration.
This soy bean proteinous soln is heated to 50 ℃, adds the 0.04 weight portion phytase catabolic enzyme " ス ミ チ-system PHY " of new Japanese chemical industry (strain) system, make it to react 60 minutes.At 150 ℃ this reactant liquor sterilization after 7 seconds, is cooled to 50 ℃, is adjusted to pH7.0, add 0.16 weight portion " プ ロ テ ア one ゼ M " of amano pharmaceutical (strain) system, make it to react 5 hours with NaOH.Be adjusted to pH6.5, after 7 seconds, use the spray dryer powder for drying immediately 150 ℃ of sterilizations.
Phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content with in this powder of vanadium molybdic acid Their Determination by Spectrophotometry does not detect (detecting boundary 5mg/100g).The mean molecule quantity result about 500 of electrophoresis technique determining.
Not (Production Example 2) (not removing the manufacturing of the soybean protein hydrolyate of phytic acid)
100 weight portions (following is " part ") soybean protein isolates (" the Off ジ プ ロ-R " of only system oil (strain) system) are made 5% aqueous solution of pH7, (big and change into (strain) system: the aspergillus origin) 1 part with プ ロ チ Application FN, at 50 ℃ of enzymolysis after 5 hours, made enzyme deactivation in 30 minutes 70 ℃ of heating, with the supernatant spray-drying that cooling back centrifugation obtains, make soybean protein hydrolyate.In addition, the TCA of this hydrolysate (trichloroacetic acid) soluble rate (the solvable nitrogen of 15%TCA/full nitrogen on duty with 100 value) is 100, and mean molecule quantity is 676.
(embodiment 1 and Comparative Examples 1)
Embodiment 1
As if move to the 300 tail 100L experimental troughs from the flatfish of preparation raising tank, prepare 5 such grooves, as experimental trough with 38 ages in days.At experimental session, the temperature of seawater remains on 17 ℃.
As the experiment feed, in the flatfish breeding feed of table 1, as shown in table 1, the low-phytate soybean protein hydrolyate of replacement of interpolation 0,2.5,5.0,10.0,20.0 weight portion caseins and above-mentioned Production Example 1 the same manufacturing is made the particulate feed with conventional method.Experiment feed composition is shown in Table 1.Unit is a weight portion.The 5th group is control group.
Since the experiment of 39 ages in days, since at 9 o'clock in the morning, per 1 hour feed, feed 8 times until 16 o'clock, at 17 o'clock, gives ア Le テ ミ ア as biological feedstuff, and every day, feed was 9 times.Forage volume is decided to be 0.31g~0.50g/ time/fish to breeding feed along with the growth of age in days, and ア Le テ ミ ア is decided to be 55~80/time/fish.
The body length of the juvenile fish seedling of about 50 tails, 52 ages in days, survive mantissa, pigment anomaly rate are measured in back 14 days of experiment beginning.
Comparative Examples 1
As the experiment feed, use and the soybean protein hydrolyate of not removing phytic acid of above-mentioned Production Example 2 the same manufacturing the method that usefulness and embodiment 1 are the same, breed flatfish.
(table 1)
Composition 1 group 2 groups 3 groups 4 groups 5 groups
The gruel of white fish fish ????30.0 ????30.0 ????30.0 ????30.0 ????30.0
Casein ????11.3 ????20.6 ????25.3 ????27.7 ????30.0
Soybean protein hydrolyate ????20.0 ????10.0 ????5.0 ????2.5 ????0.0
Dextrin ????7.0 ????7.0 ????7.0 ????7.0 ????7.0
Vitamin mixtures ????5.3 ????5.3 ????5.3 ????5.3 ????5.3
Mineral mixture ????5.0 ????5.0 ????5.0 ????5.0 ????5.0
Walleye pollack cod-liver oil ????8.0 ????8.0 ????8.0 ????8.0 ????8.0
The n-3 highly unsaturated fatty acid ????0.5 ????0.5 ????0.5 ????0.5 ????0.5
Soybean lecithin ????4.0 ????4.0 ????4.0 ????4.0 ????4.0
Cellulose ????0.9 ????1.6 ????1.9 ????2.0 ????2.2
Casein ????8.0 ????8.0 ????8.0 ????8.0 ????8.0
Add up to ????100 ????100 ????100 ????100 ????100
The result of embodiment 1 and Comparative Examples 1 is as shown in table 2.
(table 2) 52 age in days measurement results
Embodiment 1: remove phytic acid Comparative Examples 1: do not remove phytic acid
Group ????N Average body long (mm) The survival number Group ????N Average body long (mm) The survival number
????1 ????48 ????18.11 ????252 ????1 ????52 ????17.91 ????194
????2 ????54 ????17.89 ????271 ????2 ????49 ????18.18 ????215
????3 ????50 ????17.56 ????244 ????3 ????49 ????17.43 ????200
????4 ????54 ????17.36 ????230 ????4 ????54 ????17.31 ????195
????5 ????52 ????17.15 ????189 ????5 ????53 ????17.01 ????194
From the result of embodiment 1 as can be known, added 1~4 group of the low-phytate soybean protein hydrolyate and compared with in contrast 5 groups, growth all obtains promoting, does not particularly find marked difference at 1~3 group.Even for the survival number, compare, can see good tendency at 1~4 group with 5 groups.In addition, each group of pigment anomaly rate does not have difference, giving the group of soybean protein hydrolyate, does not note abnormalities yet.
In addition, compare with following comparative example 1, the survival rate of the soybean protein hydrolyate that phytic acid content is low (oligo peptide) also significantly rises.
(Production Example 3)
100 weight portion soybean protein isolates (only system oil (strain) system " ニ ュ-Off ジ プ ロ-R ") are dissolved in 900 parts in the water, toward wherein adding 2 parts of protein decomposition enzymes (big and change into Co., Ltd.'s system " プ ロ チ Application "), 50 ℃ cultivate 5 hours after, remove insoluble matter with centrifugation (5000rpm * 30 minute), further 80 ℃ of heating 30 minutes, make enzyme deactivation and sterilization, carry out freeze drying, obtain enzyme analyte (SH).
In the post of diameter 1.4cm, fill acrylic compounds weak-base anion-exchange resin (Sumitomo Chemical (strain) system " KA890 ") until height 15cm (resin volume 23cm 3), feed 50ml 5% caustic soda liquid and 500ml ion exchange water, wash.
On the other hand, dissolve above-mentioned zymolyte, regulate pH to 4.5, regulate so that final protein concentration reaches 10% with ion exchange water with 10% hydrochloric acid with ion exchange water.
Feed above-mentioned modulating liquid with 51ml/hr from the top of the post of filling KA890 and washing, from the lower part of post from the treatment fluid that obtains wash-out.
Feed enzymolysis liquid, collect its amount and finally be the enzymolysis liquid of 1219ml (as protein content is 121.9g, and every 1ml resin is 5.3g), i.e. the eluent removed by resin adsorption fully of phytic acid, freeze drying obtains the zymolyte (SHR) of low phosphorus content.Method (Cereal Chem.63.475.1986) with people such as Mohammed, founder of Islam is measured phytic acid content.Its result does not detect phytic acid (detecting boundary 0.005 weight %).
(embodiment 2 and Comparative Examples 2)
Be used in low-phytin soybean protein hydrolyate that obtains in the Production Example 3 and the soybean protein hydrolyate of not removing phytic acid that in Production Example 2, obtains, hatching is from the embryonated egg of female shrimp, raise to the feed of zoea l phase with preparation, carry out raising experiment.
The raising condition is as shown in table 3, and 100 1 phases of tail water flea larva of income at room temperature raise with the particulate feed in 1 liter of beaker.The composition of test feed is as shown in table 4.
As test feed, add 10.0 weight portion caseins (contrast) respectively in the prawn culturing feed of table 4 and replace and Production Example 3 each soybean protein hydrolyate with Production Example 2 the same manufacturings, make the particulate feed with conventional method.
In addition, judge additive effect with growth of seedlings index (Growth index: reach the growth stage) and survival rate.The result is as shown in table 5.
The raising condition of (table 3) prawn seedling
Condition Content
Seedling (when raising beginning) feeding time stocking density water temperature is raised water and is changed water feed frequency The zoea l phase reaches the preceding 100 tail/jars of postlarva, (1L) room temperature, (25~27 ℃) pH8.4 ± 0.2 100%/sky 2 times/day
Feed particle size zoea l phase mysis stage postlarva ? 53μm 125μm 250μm
Take the photograph bait rate (mg feed/seedling/sky)
1 phase of seedling 0.16
Mysis stage 0.20
Back seedling 0.24
(table 4) test feed is formed
Composition/group Contrast Production Example 3 Production Example 2
Casein ????42.8 ????32.8 ????32.8
L-arginine HCl ????2.4 ????2.4 ????2.4
The low-phytate soybean protein hydrolyate ????0.0 ????10.0 ????10.0
Glucose ????5.5 ????5.5 ????5.5
Sucrose ????10.0 ????10.0 ????10.0
Alphalise starch ????4.0 ????4.0 ????4.0
Cod liver oil ????4.0 ????4.0 ????4.0
ω3-HUFA?※1 ????0.5 ????0.5 ????0.5
Soybean lecithin ????3.0 ????3.0 ????3.0
Cholesterol ????1.0 ????1.0 ????1.0
Mineral mixture ????8.6 ????8.6 ????8.6
Vitamin mixtures ????3.0 ????3.0 ????3.0
Gucosamine HCl ????0.8 ????0.8 ????0.8
Natrium citricum ????0.3 ????0.3 ????0.3
Sodium succinate ????0.3 ????0.3 ????0.3
Antler glue ????5.0 ????5.0 ????5.0
Cellulose ????8.8 ????8.8 ????8.8
Add up to ????100.0 ????100.0 ????100.0
※1(20:5ω3)∶(22:6ω3)=3∶2
The result of the raising experiment of (table 5) prawn seedling
Group The 11st day body long (mm) Survival rate (%) Growth index (the 11st day)
Contrast ????4.56±0.27 ????79.5 ????6.9
Production Example 3 ????4.68±0.25 ????91.2 ????7.7
Production Example 2 ????4.66±0.26 ????83.5 ????7.4
(condition is growth index (Growth index)=(1n1+2n2+3n3+4n4+5n5+6n6+7n7)/N, N=n1+n2+...+n7)
N1, n2, n3, n4, n5, n6 and n7 represent the mantissa of 1 phase of zoea, 2 phases of zoea, 3 phases of zoea, 1 phase of mysis, 2 phases of mysis, 3 phases of mysis and postlarva 1 seedling
This found that, the body of postlarva long (measuring in the 11st day) does not have difference between the soybean protein hydrolyate interpolation district that the low-phytin soybean protein hydrolyate interpolation district and the nothing of Production Example 3 are added district or Production Example 2, but discovery low-phytate soybean protein hydrolyate adds the district good tendency is being arranged aspect survival rate, the growth index.
(embodiment 3)
Move on in the 20L experimental trough giving birth to the 70th~80 day the flatfish in back, each distinguishes 20 tails, prepares 3 such tanks, carries out raising experiment.Experimental session is raised water temperature and is remained on 18 ℃.
As the experiment feed, (EP feed, the seedling S-6 of (strain) ヒ ガ シ マ Le system) is modulated into base-material with commercially available flatfish feed.Just with commercially available feed heating, apply low-phytate soybean protein hydrolyate that makes manufacturing in above-mentioned Production Example 1 and the soybean protein hydrolyate of making and its adhesion without phytase treatment in Production Example 2 after, drying is made the experiment feed.The composition of commercially available feed and the composition that has added the experiment feed of soybean protein hydrolyate are shown in Table 6.
(table 6)
(composition of commercially available feed) 75.5% fish meal, rotten 12.7% wheat flour of krill, starch, corn flour 11.8% feed yeast, refined fish oil, soybean lecithin, yeast extract, calcium dihydrogen phosphate, trace element, vitamin
(adding the composition of the feed of soybean protein hydrolyate) 67.5% fish meal, rotten 10% soybean protein hydrolyate, 11.7% wheat flour of krill, starch, corn flour 10.8% feed yeast, refined fish oil, soybean lecithin, yeast extract, calcium dihydrogen phosphate, trace element, vitamin
(adding the composition of the feed of low-phytin soybean protein hydrolyate) 67.5% fish meal, rotten 10% low-phytin soybean protein hydrolyate, 11.7% wheat flour of krill, starch, corn flour 10.8% feed yeast, refined fish oil, soybean lecithin, yeast extract, calcium dihydrogen phosphate, trace element, vitamin
Begin experiment with the fry of the average 80.3mm of total length, fish body weight 4.4g, body weight/total length=55.5mg/mm, feed is since at 9 o'clock in the morning, per 6 hours feeds, and give 4 every day.Through after 10 days, 20 days, measure long, the fish body weight of body of juvenile fish seedling 60 tails after the experiment beginning, calculate body weight/total length from measurement result.
(table 7) measures beginning the 0th day (several 60 tails of test fish)
The check plot Soybean protein hydrolyate Low phytin soybean protein hydrolyate
Long (mm) minimax of average body ????81.4 ????90 ????73 79.0 88 72 ????80.4 ????90 ????71
Average weight (g) minimax ????4.49 ????6.5 ????2.9 4.26 6.2 3.1 ????4.54 ????6.0 ????3.1
Long (mg/mm) minimax of average weight/body ????55.2 ????72.2 ????39.7 53.9(-1.3) 70.5(-1.7) 43.1(+3.4) ????56.5(+1.3) ????66.7(-5.5) ????43.7(+4.0)
Remarks: () interior increase and decrease with respect to the check plot
(table 8) measures beginning the 10th day (several 60 tails of test fish, survival rate 60 tails)
The check plot Soybean protein hydrolyate Low phytin soybean protein hydrolyate
Long (mm) minimax of average body ????86.1 ????95 ????73 ????82.5 ????95 ????74 ????86.4 ????101 ????71
Average weight (g) minimax ????4.99 ????6.8 ????2.7 ????4.57 ????7.3 ????4.1 ????5.31 ????8.2 ????3.2
Long (mg/mm) minimax of average weight/body ????57.9 ????71.6 ????37.0 ????55.4(-2.5) ????76.8(+5.2) ????43.2(+6.2) ????61.5(+3.6) ????81.2(+9.6) ????53.9(+16.9)
Remarks: () interior increase and decrease with respect to the check plot
(table 9) measures beginning the 20th day (several 60 tails of test fish, survival rate 60 tails)
The check plot Soybean protein hydrolyate Low phytin soybean protein hydrolyate
Long (mm) minimax of average body ????94.0 ????106 ????76 ????89.3 ????106 ????78 ????93.4 ????101 ????81
Average weight (g) minimax ????7.02 ????10.6 ????3.3 ????6.05 ????10.9 ????3.9 ????6.97 ????11.8 ????5.3
Long (mg/mm) minimax of average weight/body ????74.7 ????100.0 ????43.4 ????67.7(-10.0) ????102.8(+2.8) ????50.0(+6.6) ????74.6(-0.1) ????106.3(+6.3) ????65.4(+22.0)
Remarks: () interior increase and decrease with respect to the check plot
Found that from this, compare, add the district,,, have the effect that promotion is grown in the growth rate height of average weight/body long (mg/mm) at the 10th day at the low-phytin soybean protein hydrolyate with control group district, interpolation soybean protein hydrolyate.Further,, also observe maximum solid for body weight/body long (mg/mm) at the 20th day.Minimum compares with check plot, interpolation soybean protein hydrolyate, and value is also than higher, and the chances are for this because feed concentrates in the big fish of solid, and the difference between solid is big, the not too big difference of mean value and check plot.
(feeding process)
Water temperature between feeding period, feeding coal, picked-up situation are described below.
The situation of ight soil, the situation of the ight soil behind the feed of check plot is for having loose bowels just the water muddiness of tank, in contrast, add the district at soybean protein hydrolyate and the big calcium magnesium of low-phytate legumin hydrolysate, ight soil is the soft stool shape of maintenance shape, and the water of tank keeps clean.And there is the ight soil thickness in the feed of low-phytin soybean protein hydrolyate, the tendency that ight soil is many.
(table 10)
The day moon Number of days Water temperature Target area Soybean protein hydrolyate The low-phytate soybean protein hydrolyate
Feeding coal The picked-up situation The survival number Feeding coal The picked-up situation The survival number Feeding coal The picked-up situation The survival number
??4/2 ??1 ????17.7 ????0 ??20 ??0 ??20 ??0 ??20
??4/3 ??2 ????18.0 ????8 Torpescence, but picked-up ??20 ??8 Torpescence, but picked-up ??20 ??8 Torpescence, but picked-up ??20
??4/4 ??3 ????18.8 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy, absorb ??20
??4/5 ??4 ????18.3 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy picked-up well ??20
??4/6 ??5 ????18.3 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy picked-up well ??20
??4/7 ??6 ????18.4 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy, absorb ??20
??4/8 ??7 ????18.4 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy, absorb ??20
??4/9 ??8 ????18.4 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy, absorb ??20
??4/10 ??9 ????17.8 ????4 The water muddiness is absorbed ??20 ??4 Not muddy, absorb ??20 ??4 Not muddy, absorb ??20
??4/11 ??10 ????18.3 ????5 The water muddiness is absorbed ??20 ??5 Not muddy, absorb ??20 ??5 Not muddy, absorb ??20
??4/12 ??11 ????18.5 ????5 The water muddiness is absorbed ??20 ??5 Not muddy, absorb ??20 ??5 Not muddy, absorb ??20
??4/13 ??12 ????18.5 ????5 The water muddiness is absorbed ??20 ??5 Not muddy, absorb ??20 ??5 Not muddy picked-up well ??20
??4/14 ??13 ????18.6 ????5 The water muddiness is absorbed ??20 ??5 Not muddy, absorb ??20 ??5 Not muddy, absorb ??20
??4/15 ??14 ????18.8 ????5 The water muddiness is absorbed ??20 ??5 Not muddy, absorb ??20 ??5 Not muddy, absorb ??20
??4/16 ??15 ????18.9 ????8 The water muddiness is taken the photograph ??20 ??8 It is not muddy, ??20 ??8 It is not muddy, ??20
Get Absorb Absorb
??4/17 ??16 ????19.4 ????6 The water muddiness is absorbed ??20 ??6 Not muddy, absorb ??20 ??6 Not muddy, absorb ??20
??4/18 ??17 ????18.4 ????6 The water muddiness is absorbed ??20 ??6 Not muddy, absorb bad ??20 ??6 Not muddy, absorb bad ??20
??4/19 ??18 ????18.4 ????6 The water muddiness is absorbed ??20 ??6 Not muddy, absorb bad ??20 ??6 Not muddy, absorb bad ??20
??4/20 ??19 ????18.4 ????6 The water muddiness is absorbed ??20 ??6 Not muddy, absorb bad ??20 ??6 Not muddy, absorb bad ??20
??4/21 ??20 ????18.4 ????8 The water muddiness is absorbed ??20 ??8 Not muddy, absorb ??20 ??8 Not muddy, absorb ??20
The following describes the embodiment of low-phytate soybean protein hydrolyate.
(embodiment 4)
In the 10kg defatted soybean, add 7 times water, under 50 ℃, pH7, stir to extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, with sulfuric acid soya-bean milk is adjusted to pH4.5 after, by centrifugation etc., after being separated into isoelectric point precipitates albumen matter and whey protein, with respect to isoelectric point precipitates albumen matter, add 4 times of water after, be adjusted to pH7.0 with NaOH, the soy bean proteinous soln of 50 liter of 8% concentration of preparation.
In this soy bean proteinous soln, add " プ ロ テ ア one ゼ S " and " プ ロ テ ア one ゼ M " (amano pharmaceutical (strain) system) each 160g respectively, make it to react 5 hours, add water decomposition (15%TCA soluble rate 85%) at 50 ℃.Then, pH is adjusted to 6.0, adds the 40g phytic acid catabolic enzyme " ス ミ チ-system PHY " of new Japanese chemical industry (strain) system, 50 ℃ make it to react 60 minutes after, be adjusted to 6.5 with NaOH, after 7 seconds, use the spray dryer powder for drying immediately 150 ℃ of sterilizations.
Phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content with in this powder of vanadium molybdic acid Their Determination by Spectrophotometry does not detect (detecting boundary 5mg/100g), admirably the sinking low-phytate.
(embodiment 5)
In the 10kg defatted soybean, add 7 times water, under 50 ℃, pH7, stir to extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, with sulfuric acid soya-bean milk is adjusted to pH4.5 after, by centrifugation etc., after being separated into isoelectric point precipitates albumen matter and whey protein, with respect to isoelectric point precipitates albumen matter, add 4 times of water after, be adjusted to pH6.0 with NaOH, the soy bean proteinous soln of 50 liter of 8% concentration of preparation.
This soy bean proteinous soln is heated to 50 ℃, add the phytase catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 40g (strain) system, after making it to react 60 minutes, be adjusted to 7.0 with NaOH, add " the プ ロ テ ア one ゼ M " of 160g amano pharmaceutical (strain) system, make it to react 5 hours.Be adjusted to pH6.5, after 7 seconds, use the spray dryer powder for drying immediately 150 ℃ of sterilizations.
Phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content with in this powder of vanadium molybdic acid Their Determination by Spectrophotometry does not detect (detecting boundary 5mg/100g), admirably the sinking low-phytate.
(embodiment 6)
In the 10kg defatted soybean, add 7 times water, under 50 ℃, pH7, stir and extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, after with sulfuric acid soya-bean milk being adjusted to pH4.5, by centrifugation etc., be separated into isoelectric point precipitates albumen matter and whey protein after, with respect to isoelectric point precipitates albumen matter, after adding 4 times of water, be adjusted to pH6.0 with NaOH, the soy bean proteinous soln of 50 liter of 8% concentration of preparation is used the spray dryer powder for drying immediately.
This protein liquid is adjusted to 8% solution, be heated to 50 ℃, add the phytase catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 40g (strain) system, after making it to react 60 minutes, be adjusted to 7.0 with NaOH, add " the プ ロ テ ア one ゼ M " of 160g amano pharmaceutical (strain) system, make it to react 5 hours.Be adjusted to pH6.5, after 7 seconds, use the spray dryer powder for drying immediately 150 ℃ of sterilizations.
With phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content in this powder of vanadium molybdic acid Their Determination by Spectrophotometry, phytic acid is reduced to 0.5% (detecting boundary 5mg/100g).
(embodiment 7)
In the 10kg defatted soybean, add 7 times water, under 50 ℃, pH7, stir to extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, with sulfuric acid soya-bean milk is adjusted to pH4.5 after, by centrifugation etc., after being separated into isoelectric point precipitates albumen matter and whey protein, with respect to isoelectric point precipitates albumen matter, add 4 times of water after, be adjusted to pH6.0 with NaOH, the soy bean proteinous soln of 50 liter of 8% concentration of preparation.
This soy bean proteinous soln is heated to 50 ℃, adds the phytase catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 40g (strain) system, make it to react 60 minutes after, use the spray dryer powder for drying immediately.
This solution is adjusted to 8% solution, is adjusted to 7.0, add " the プ ロ テ ア one ゼ M " of 160g amano pharmaceutical (strain) system, make it to react 5 hours, after 7 seconds, use the spray dryer powder for drying immediately 150 ℃ of sterilizations with NaOH.
With phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content in this powder of vanadium molybdic acid Their Determination by Spectrophotometry, phytic acid is reduced to 0.2% (detecting boundary 5mg/100g).
(embodiment 8)
In the 10kg defatted soybean, add 7 times water, under 50 ℃, pH7, stir to extract after 30 minutes, separate soybean residue and soya-bean milk with centrifugal separator, with sulfuric acid soya-bean milk is adjusted to pH4.5 after, by centrifugation etc., after being separated into isoelectric point precipitates albumen matter and whey protein, with respect to isoelectric point precipitates albumen matter, add 4 times of water after, be adjusted to pH6.0 with NaOH, the soy bean proteinous soln of 50 liter of 8% concentration of preparation.
This protein liquid is heated to 50 ℃, adds the phytase catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 40g (strain) system, make it to react 60 minutes after, use the spray dryer powder for drying immediately.
This solution is adjusted to 8% solution, be adjusted to 7.0 with NaOH, add " the プ ロ テ ア one ゼ M " of 160g amano pharmaceutical (strain) system, make it to react 5 hours (pH6.2 during reaction), afterwards, add the phytic acid catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 40g (strain) system again, make it to react 60 minutes after, after 7 seconds, use the spray dryer powder for drying 150 ℃ of sterilizations immediately.
With phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content in this powder of vanadium molybdic acid Their Determination by Spectrophotometry, phytic acid is reduced to 0.2% (detecting boundary 5mg/100g).
(embodiment 9)
The soybean protein isolate of 30kg powder for drying (only system oil (strain) system, " ニ ュ-Off ジ プ ロ-R ") make 10% solution of pH7.0, make it to be subjected to the effect of protease " プ ロ テ ア one ゼ S " and " the プ ロ テ ア one ゼ M " that 0.3kg amano pharmaceutical (strain) is made of 1.2kg amano pharmaceutical (strain) system, after 50 ℃ of hydrolysis 5 hours (15%TCA soluble rate 85%), pH is adjusted to 6.0, add the phytase catabolic enzyme " ス ミ チ-system PHY " of the new Japanese chemical industry of 0.6kg (strain) system, 45 ℃ of hydrolysis 2 hours.
In can processed continuously high-speed centrifuge, this decomposed solution is adjusted to 100 liters/hour liquor charging speed, separate and remove the precipitate component of generation.Gained centrifuged supernatant (yield of solid content is 70%) is adjusted to pH6.5, after 7 seconds, uses the spray dryer powder for drying immediately 150 ℃ of sterilizations.
Phytic acid (meso inositol cyclohexylenedinitrilotetraacetic acid) content with in this powder of vanadium molybdic acid Their Determination by Spectrophotometry does not detect (detecting boundary 5mg/100g), admirably the sinking low-phytate.
                       Industrial applicibility
By with feed feed of the present invention, can promote growing of juvenile fish seedling, increase substantially survival rate.
Particularly can improve the survival rate of juvenile fish seedling that it is generally acknowledged the where the shoe pinches of breeding fish, can cultivate fish that in the past it is generally acknowledged the difficulty of breeding fish.
In addition, can promote growing of the fry that grows up to a certain extent, can make the ight soil thickness, anti-sealing is muddy, and growing environment is improved.
Also have, it is possible that hydrolyzate of low-phytin vegetable protein is suitable for this number of animals raised.
That is to say that according to the present invention, phytic acid content is few, and to detect with vanadium molybdic acid absorption photometry be that to detect the following hydrolyzate of low-phytin vegetable protein of boundary be possible. In addition, compare the local flavor of the hydrolyzate of low-phytin vegetable protein of the inventive method excellent (being because the analyte of phytic acid by inference) with the low-phytin soybean protein hydrolyate of resin adsorption method.
In addition, the molecular weight of hydrolyzate of low-phytin vegetable protein of the present invention is little, phytic acid is few, therefore be used for the less developed fry of digestion and absorption function and give birth to after survive animal feed the time, the property digested and assimilated excellence is extremely effective to the absorption that promotes the trace meters such as calcium.

Claims (10)

1. a feed for fry young fishes is characterized in that containing the vegetable protein hydrolyzate that phytic acid content is 0.05 weight % or following (dry solid content meter) in feedstuff.
2. the described feed for fry young fishes of claim 1, wherein, described vegetable protein hydrolyzate is that mean molecule quantity is 200~10000 vegetable protein hydrolyzate.
3. claim 1 or 2 described feed for fry young fishes, wherein, the form of described feed is with the particle diameter that keeps juvenile fish to be easy to absorb, floatability, sinking speed and not stripping of nutrient in water, the particulate feed of the microencapsulation of being digested and assimilated in alimentary canal.
4. the manufacture method of a hydrolyzate of low-phytin vegetable protein is characterized in that, this method comprises the operation that (a) decomposes phytic acid with the operation of protease decomposing protein and the enzyme of (b) using the decomposition phytic acid.
5. the described manufacture method of claim 4 wherein, behind the protease decomposing soya-bean protein, is decomposed phytic acid with the enzyme that decomposes phytic acid.
6. the described manufacture method of claim 4, wherein, decompose undried soybean protein with the enzyme that decomposes phytic acid after, use the protease decomposing protein.
7. any described manufacture method in the claim 4~6, wherein, the enzyme of described decomposition phytic acid is a phytase.
8. any described manufacture method in the claim 4~7, wherein, the pH of described phytase treatment is 6~9.
9. any described manufacture method in the claim 4~8, wherein, the mean molecule quantity of described hydrolyzate of low-phytin vegetable protein is 200~10000.
10. any described manufacture method in the claim 4~9, wherein, phytic acid content in dry solid content in the described low-phytate vegetable protein hydrolyzate is shown as, and can not detect phytic acid with vanadium molybdic acid absorption photometry (detecting boundary 5mg/100g).
CNB038177072A 2002-07-24 2003-07-23 Feed for fry young fishes and method of producing hydrolyzate of low-phytin vegetable protein to be used therein Expired - Fee Related CN100379354C (en)

Applications Claiming Priority (6)

Application Number Priority Date Filing Date Title
JP2002215660 2002-07-24
JP215660/2002 2002-07-24
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