WO1997000020A1 - Compose pour les animaux ou l'homme a prendre par voie orale et contenant des enzymes, et procede pour produire ledit compose - Google Patents

Compose pour les animaux ou l'homme a prendre par voie orale et contenant des enzymes, et procede pour produire ledit compose Download PDF

Info

Publication number
WO1997000020A1
WO1997000020A1 PCT/JP1996/001579 JP9601579W WO9700020A1 WO 1997000020 A1 WO1997000020 A1 WO 1997000020A1 JP 9601579 W JP9601579 W JP 9601579W WO 9700020 A1 WO9700020 A1 WO 9700020A1
Authority
WO
WIPO (PCT)
Prior art keywords
enzyme
accession number
composition
ferm
bacillus
Prior art date
Application number
PCT/JP1996/001579
Other languages
English (en)
Japanese (ja)
Inventor
Shinobu Ito
Motoaki Kamachi
Hajime Sato
Original Assignee
Novo Nordisk A/S
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Novo Nordisk A/S filed Critical Novo Nordisk A/S
Publication of WO1997000020A1 publication Critical patent/WO1997000020A1/fr

Links

Classifications

    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes

Definitions

  • the present invention has an effect that it is not decomposed even in the process of producing a molded composition using a high-temperature heating type device, and is effective against a wide range of domestic animals such as livestock, poultry, and pet farmed animals.
  • a thermostable enzyme the effects derived from its enzyme activity can be achieved in a wide variety of animal species. It has been confirmed that the present invention is sufficiently exhibited, and the present invention has been completed.
  • composition for oral use of animals according to 1) above which is for feed, food or medicine.
  • This enzyme has a residual activity of 60% or more when heated from a culture solution in a constant temperature bath at 80 ° C for 20 seconds.
  • the carbon source may be any assimilable carbon compound or a compound containing the same.
  • glucose, maltose, starch, carboxymethylcellulose (CMC) and the like can be used.
  • Polysaccharides assimilated with enzymes produced by bacteria may be used in combination.
  • xylanase various xylan, wheat bran, pulp waste liquor, xylan such as cereal saccharified kashiwa or rice straw, or various raw materials containing xylan-based polysaccharide, or starch hydrolyzation such as glucose, starch or liquefied starch
  • sugars such as molasses can be used alone or in combination.
  • the mutant gene may be integrated into the chromosome or replicated in the host cell.
  • E. coli plasmid is useful in the former case.
  • Bacillus subtilis vectors JP-A-61-162187, etc.
  • the host cell may be any as long as it can introduce the modified enzyme gene and can express and produce it, but preferably secretes the produced enzyme outside the cells.
  • protease Bacillus bacterium has such properties, so that Bacillus bacterium, for example, NKS-21 described above can be used.
  • compositions for oral use of animals of the present invention include various terrestrial animals, for example, livestock such as pigs, cattle and horses, poultry such as chickens, dogs such as dogs and cats, and various marine animals.
  • livestock such as pigs, cattle and horses
  • poultry such as chickens
  • dogs such as dogs and cats
  • various marine animals for example, fish, such as salmon, hamachi, trout, trout, carp, eel, shrimp, eel, crustaceans, etc.
  • various insects such as silkworms, which are fed to animals for oral growth It is a feed composition.
  • substances other than enzymes can be the same as those conventionally used in accordance with the animal species to be orally administered.
  • the effect of the enzyme can be further enhanced by using a substance containing a large amount of components that serve as substrates for the enzyme. It may also contain useful bacteria having a function of decomposing or converting feed crops into useful forms, such as lactic acid bacteria in silage processing.
  • the composition for oral administration to animals of the present invention can be produced according to a method known per se, but a heating-type device is used in the production process because an enzyme having higher heat resistance is used than before.
  • a big feature is what you can do. That is, after the above-mentioned various nutrients are mixed in a predetermined composition ratio, the enzyme of the present invention is added to the obtained mixture, crushed and mixed, and then, if necessary, molded.
  • the enzyme may be added in either powder or liquid form. The added enzyme mixes well with other components. Use enzymes in both powder and liquid If necessary, the processing efficiency can be increased by using diluents, diluents, and concentrates to ensure good mixing.
  • heating-type equipment examples include a pellet mill, a pelletizer machine, an extruder, an extruder, and the like, which can be generally used to pelletize feed.
  • Fluidized bed granulators, centrifugal rolling granulators, floating fluid granulators, swirling fluid granulators, etc. can also be used as appropriate according to the intended use.
  • the effect becomes more remarkable when a heat-press type molding machine (for example, an extruder) is used.
  • a heat-press type molding machine for example, an extruder
  • the use of such equipment makes it possible to efficiently gelatinize the starch in the raw material composition and thus excel in pellet moldability.
  • the ability to add a stable enzyme during such a molding process is a function of the product. This is extremely useful for improving cost performance and reducing costs by preventing enzyme deactivation.
  • the heat-resistant enzymes according to the present invention have high stability even at a high temperature of 70 ° C. or more, it can be said that they are extremely advantageous in producing an animal oral composition through such a molding step.
  • This enzyme-containing solution was suspended in 100 times the volume of distilled water at 80 ° C and the remaining activity was measured for 20 seconds in a thermostat at 80 ° C.
  • the enzyme activity was 1.84SU. / m 1 (residual rate 92%).
  • Comparative Example 1 Production of enzyme using Bacillus sp. NK S-21
  • the enzymatic activity of the NKS-21 culture medium was 85 nkat Zm1. This enzyme solution was suspended in 100 times the volume of distilled water at 80 ° C and the remaining activity was measured for 20 seconds in a constant temperature bath at 80 ° C. The enzyme activity was 0.07 nkat / ml (residual rate 8%).
  • Example 4 Enzyme production using Bacillus sp. NKS-21 mutant
  • Plasmid pSDT812 Japanese Patent Application Laid-Open No. 1-1415966 in which a wild-type al force reprotease gene cloned from Bacillus NKS-21 has been inserted into the Clal cleavage site of pHSG396.
  • EcoRI and BamHI also digested with the restriction enzymes EcoRI and BamHI, mixed with pUC118 that had been treated with alfa phosphatase, and the T4 phage DN
  • Escherichia coli JM101 was transformed with this DNA, and transformants resistant to ampicillin and sensitive to kanamycin were selected.
  • Plasmid DNA was extracted, purified, and analyzed by a conventional method using these transformants. As a result, the wild-type plasmid of pSDT812 was located between the EcoRKBamHI cleavage points of pUC118.
  • the plasmid pSD3001 into which the EcoRI—BamHI fragment containing the gene for the lipoliporase was inserted was obtained.
  • This plasmid pSD3001 contains a gene having the nucleotide sequence of SEQ ID NO: 1.
  • the site-specific mutation was performed according to the Kunkel method as follows.
  • Plasmid p SD301 was transformed into E. coli BW313 (H fr KL16P0 / 45 [1ysA (61-62)], dutl, nug1, thy—1, re1A). 1) to obtain Escherichia coli BW313 having the plasmid pSD301. From this E. coli, the method of Messing et al. (Vielra, J. and
  • the Origonukureochi ⁇ twenty-one-ring the single-stranded DNA described above using de carbonochloridate Ffa (2 0 mM T ris' HC l H 8.0, 1 0 mM M g C 1 2, 5 0 mM N a C 1, (1 mM DTT), left at 65 ° C for 15 minutes, and then left at 37 ° C for 15 minutes to allow oligonucleotides to anneal to the target site of the mutation.
  • de carbonochloridate Ffa 2 0 mM T ris' HC l H 8.0, 1 0 mM M g C 1 2, 5 0 mM N a C 1, (1 mM DTT
  • a polymerase was added, and the mixture was allowed to stand at 25 ° C for 2 hours to synthesize a complementary strand. Thereafter, this DNA was transformed into Escherichia coli BMH71-18mutS according to a conventional method, and an ampicillin-resistant transformant was selected. From some of the obtained transformants, single-stranded DNA was prepared according to a conventional method, and the nucleotide sequence was determined by the dideoxy method.A plasmid corresponding to Ser12A1a was generated. SD 3002 was obtained. pSD3002 was digested with restriction enzymes E.coRI and BamHI, mixed with pUBllO similarly digested with EcoRI and BamHI, and treated with T4 DNA ligase.
  • Bacillus subtilis BD224 was transformed by the protoplast method. Kanamycin-resistant transformants were selected that produced a large clear zone on an agar medium containing 2% skim milk. Plasmid DNA was extracted and purified by a conventional method using these transformants. Plasmid pSD3102 in which the EcoRI—BamHI fragment containing the gene for pSD3002 was inserted between the 10 EcoRI—BamHI cleavage sites was inserted. Obtained.
  • Bacillus NKS-21 was transformed by the protoplast method, and kanamycin-resistant transformation was selected, whereby serine at position 12 of the wild-type protease was replaced with alanine.
  • a mutant having the gene was obtained. This variant is abbreviated as “S12 ⁇ A” in the following description and tables.
  • a liquid medium consisting of 2.0% soybean meal, 3.5% maltose, 0.5% dihydrogen phosphate, 0.05% magnesium sulfate, and 1.0% sodium carbonate was added in two 5 ⁇ volumes. It was placed in a jar fermenter and sterilized at 121 ° C for 20 minutes. The Bacillus sp.
  • Example 2 Approximately 56% fish meal, 6% salmon milt, 5% krill meal, 5% squid viscera, 5% squid liver oil, 4% soybean lecithin, 0.5% amino acid mix, 5% corn starch, 5% mineral mix, 5 gluten 3% of the enzyme solution of Bacillus sp. SD 771 obtained in Example 1 was added to a total of 100% of the dried raw material, which was made up of 100% with 2.8%, vitamin 2.8% and magnesium 0.01%. After pulverizing this raw material and mixing it sufficiently with a mixer, put it in a pellet mill (manufactured by California Pellet Mill Co., Ltd.), raise the temperature while blowing steam, and raise the temperature in the pellet mill to 80-85.
  • a pellet mill manufactured by California Pellet Mill Co., Ltd.
  • the pellet was formed at a temperature of ° C.
  • the transit time at this time was less than 20 seconds. After that, it was dried by blowing dry air at 70 ° C for 2 hours to produce a feed for marine animals. In this feed, the starch component was pregelatinized, and the mold and hardness were good. When the enzyme activity of the amylase was measured, 95% of the enzyme activity at the time of addition remained.
  • amylase activity was determined by using soluble starch as a substrate, reacting in a 0.05 M CHES buffer ( ⁇ ⁇ 9.0) at 50 ° C, developing the color with an iodine-potassium iodide solution, and measuring the absorbance. Decided from the decrease.
  • Example 6 Production of amylase-containing feed for aquatic animals
  • Soybean flour is added to 35% fish meal, 30% corn meal, 3% MacCallum salt, 1.4% vitamin mix, 0.01% magnesium L-ascorbic acid-phosphate and 100% by adding soy flour.
  • 3% of the enzyme solution of Bacillus sp. SD771 obtained in Example 1 was added. After thoroughly mixing with a sachet, the mixture was molded by an extruder. This molded product was dried at 100 ° C. for 1 hour by a dryer to produce a pellet feed for aquatic animals. When the enzyme activity of amylase was measured in the same manner as in Example 5, it was found that 82% of the added enzyme activity remained based on the added enzyme activity.
  • Example 7 Production of xylanase-containing terrestrial animal feed
  • Rice straw having a dry weight of 500 g was cut into a length of 5 cm (sample 1) or ground with a mixer (sample 2), immersed in water 33, and left at room temperature for 24 hours. After that, the rice straw is taken out, washed thoroughly with water to remove the eluate, mud and other deposits, centrifugally dehydrated, and then water is added so that the raw material becomes 15%, pH 7.0. The temperature was adjusted to 70 ° C. To this, the enzyme solution derived from Bacillus sp. SD902 obtained in Example 2 was gently added so as to be 0.2% (vs. dry rice straw) and stirred for 24 hours. . After the reaction, the rice straw was crushed and washed with water, and then centrifuged and dehydrated.
  • the lignin content of the treated rice straw was reduced by 18% for sample 1 and 25% for sample 2, indicating that the rice straw had changed to a digestible rice straw (or ground product). That is, it was confirmed that the xylanase-containing solution obtained in Example 2 had useful properties leading to improvement in degradability.
  • Table 2 shows the results of similar experiments conducted with various enzyme solutions obtained in Examples 1 to 4 and a combination of these two enzyme solutions, together with the results obtained with the enzyme solution obtained in Comparative Example 1. Show. Those with an effect were marked with ⁇ (especially marked with ⁇ ), and those with no effect were marked with X. As shown in these results, the heat-resistant enzyme-containing feed of the present invention shows a remarkable effect of improving digestibility.
  • various enzymes can be used in the present invention, they are useful as feed compositions for a wide range of organisms such as livestock, pets, poultry, marine animals, insects, and the like.
  • starch can be pregelatinized in the heat molding process of manufacturing enzyme-compounded molded articles, so that Can be efficiently produced.
  • the technology of the present invention can be used in accordance with the method specifically described for the feed composition herein, for example, ingestion through the mouth of animals including humans, fish, birds, etc., such as foodstuffs and drugs. Widely used in animal oral compositions.
  • Sequence type nucleic acid

Abstract

L'invention concerne un composé à prendre par voie orale destiné aux animaux et qui contient des enzymes thermostables ayant une activité résiduelle de 60 % ou plus après avoir été chauffés à 80° pendant 20 secondes. Ce composé est utile en tant que pâture, aliment ou médicament qui peuvent être pris par voie orale par l'homme et par les animaux, y compris les poissons et les oiseaux, et qui sont améliorés dans leur fonctionnalité, dans leur valeur nutritive, dans leur aptitude au stockage et dans leur digestibilité in vivo.
PCT/JP1996/001579 1995-06-14 1996-06-11 Compose pour les animaux ou l'homme a prendre par voie orale et contenant des enzymes, et procede pour produire ledit compose WO1997000020A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP7/147672 1995-06-14
JP07147672A JP2001057852A (ja) 1995-06-14 1995-06-14 耐熱酵素含有飼料用組成物

Publications (1)

Publication Number Publication Date
WO1997000020A1 true WO1997000020A1 (fr) 1997-01-03

Family

ID=15435674

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP1996/001579 WO1997000020A1 (fr) 1995-06-14 1996-06-11 Compose pour les animaux ou l'homme a prendre par voie orale et contenant des enzymes, et procede pour produire ledit compose

Country Status (2)

Country Link
JP (1) JP2001057852A (fr)
WO (1) WO1997000020A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002239573A (ja) * 2001-02-21 2002-08-27 Sanyuu:Kk 水の浄化方法
JP2002281909A (ja) * 2001-03-26 2002-10-02 Oriental Yeast Co Ltd 養魚用配合飼料
JP2005535339A (ja) * 2002-08-14 2005-11-24 ズーライフ インターナショナル リミテッド 食餌の栄養強化用組成物
JP2006520601A (ja) * 2003-03-18 2006-09-14 ネステク ソシエテ アノニム 動物の健康を増進するための方法及び組成物
JP2009519712A (ja) * 2005-12-15 2009-05-21 ケムゲン コーポレーション 免疫学的ストレス低減のための酵素
RU2777100C1 (ru) * 2021-04-02 2022-08-01 Федеральное государственное бюджетное образовательное учреждение высшего образования "Уральский государственный аграрный университет" (ФГБОУ ВО Уральский ГАУ) Способ кормления яичных кур

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1471799A2 (fr) * 2002-01-25 2004-11-03 DSM IP Assets B.V. Compositions enzymatiques thermostables
GB201011513D0 (en) * 2010-07-08 2010-08-25 Danisco Method
JP6426397B2 (ja) * 2014-08-12 2018-11-21 石川県公立大学法人 アカテガニ消化管由来バイオマス分解細菌群

Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5545314A (en) * 1978-09-25 1980-03-31 Sanwa:Kk Feed additive for raising fish
JPH02215381A (ja) * 1989-02-15 1990-08-28 Showa Denko Kk 新規セルラーゼ及びその製造法
JPH0423983A (ja) * 1990-05-18 1992-01-28 Showa Denko Kk アミラーゼ及びその製造法
JPH0458885A (ja) * 1990-06-28 1992-02-25 Showa Denko Kk アミラーゼ及びその製造法
JPH0479882A (ja) * 1990-07-24 1992-03-13 Showa Denko Kk セルラーゼ及びその製造法
JPH04104788A (ja) * 1990-08-24 1992-04-07 Showa Denko Kk セルラーゼ及びその製造法
JPH0591876A (ja) * 1991-10-02 1993-04-16 Showa Denko Kk プロテアーゼ、その製造法および用途
JPH05209385A (ja) * 1992-01-29 1993-08-20 Showa Denko Kk パルプの漂白方法
JPH0626175A (ja) * 1992-07-09 1994-02-01 Sumi Kenzaiten:Kk 建築用板材の取付け方法と、建築用板材の取付け用釣子
JPH06225706A (ja) * 1992-12-10 1994-08-16 Reiko Kosaka 動物用飼料
JPH06319539A (ja) * 1993-04-05 1994-11-22 Aveve Nv フィチン酸加水分解用酵素組成物及びフィチン酸の加水分解方法

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS5545314A (en) * 1978-09-25 1980-03-31 Sanwa:Kk Feed additive for raising fish
JPH02215381A (ja) * 1989-02-15 1990-08-28 Showa Denko Kk 新規セルラーゼ及びその製造法
JPH0423983A (ja) * 1990-05-18 1992-01-28 Showa Denko Kk アミラーゼ及びその製造法
JPH0458885A (ja) * 1990-06-28 1992-02-25 Showa Denko Kk アミラーゼ及びその製造法
JPH0479882A (ja) * 1990-07-24 1992-03-13 Showa Denko Kk セルラーゼ及びその製造法
JPH04104788A (ja) * 1990-08-24 1992-04-07 Showa Denko Kk セルラーゼ及びその製造法
JPH0591876A (ja) * 1991-10-02 1993-04-16 Showa Denko Kk プロテアーゼ、その製造法および用途
JPH05209385A (ja) * 1992-01-29 1993-08-20 Showa Denko Kk パルプの漂白方法
JPH0626175A (ja) * 1992-07-09 1994-02-01 Sumi Kenzaiten:Kk 建築用板材の取付け方法と、建築用板材の取付け用釣子
JPH06225706A (ja) * 1992-12-10 1994-08-16 Reiko Kosaka 動物用飼料
JPH06319539A (ja) * 1993-04-05 1994-11-22 Aveve Nv フィチン酸加水分解用酵素組成物及びフィチン酸の加水分解方法

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2002239573A (ja) * 2001-02-21 2002-08-27 Sanyuu:Kk 水の浄化方法
JP2002281909A (ja) * 2001-03-26 2002-10-02 Oriental Yeast Co Ltd 養魚用配合飼料
JP2005535339A (ja) * 2002-08-14 2005-11-24 ズーライフ インターナショナル リミテッド 食餌の栄養強化用組成物
JP2006520601A (ja) * 2003-03-18 2006-09-14 ネステク ソシエテ アノニム 動物の健康を増進するための方法及び組成物
JP2009519712A (ja) * 2005-12-15 2009-05-21 ケムゲン コーポレーション 免疫学的ストレス低減のための酵素
RU2777100C1 (ru) * 2021-04-02 2022-08-01 Федеральное государственное бюджетное образовательное учреждение высшего образования "Уральский государственный аграрный университет" (ФГБОУ ВО Уральский ГАУ) Способ кормления яичных кур

Also Published As

Publication number Publication date
JP2001057852A (ja) 2001-03-06

Similar Documents

Publication Publication Date Title
CA2292953C (fr) Compositions de phytase a haute activite
CN100475051C (zh) 含有肌醇六磷酸酶的动物食物和方法
JP3105920B2 (ja) 細菌性フィターゼをコードする配列、及びフィターゼ活性を有するタンパク質
CN100406561C (zh) 微生物表达的用于动物饲料的耐热植酸酶
CA2120265C (fr) Hydrolyse du phytate et composition enzymatique utilisee a cette fin
RU2227159C2 (ru) Фитаза из bacillus subtilis, ген, кодирующий эту фитазу, способ ее получения и применение
JP2780833B2 (ja) ケラチン質原料の減成方法及びこれに有用な細菌
RU99105347A (ru) Фитаза из bacillus subtilus, ген, кодирующий эту фитазу, способ ее получения и применение
CN1561391A (zh) 重组细菌肌醇六磷酸酶及其用途
KR20210149775A (ko) 동물 사료 조성물에서의 산화환원 효소
EP2428565B1 (fr) Polypeptide à activité phytase et à stabilité thermique accrue de l'activité enzymatique et séquence nucléotidique codant ce polypeptide
MX2007004524A (es) Polipeptido que tiene actividad de fitasa y secuencia nucleotida que lo codifica.
CN112654703A (zh) 用于基于谷类的动物饲料的含木聚糖酶饲料添加剂
CN1188803A (zh) 使用谷物种子生产酶制品及饲料原料
WO1997000020A1 (fr) Compose pour les animaux ou l'homme a prendre par voie orale et contenant des enzymes, et procede pour produire ledit compose
US6162473A (en) Hemicellulase use in feeds with low caloric content
US20020102329A1 (en) Methods for high-temperature hydrolysis of galactose-containing oligosaccharides in complex mixtures
WO2022074170A1 (fr) Conservation enzymatique de probiotiques dans des aliments pour animaux
FI81484C (fi) Metod foer behandling av foderraovara, foderraovara och foderblandning.
CA2253806C (fr) Utilisation d'hemicellulase dans des aliments basses calories
ZA200403981B (en) Phytases and method for producing these phytases.
TWI759803B (zh) 新穎絲胺酸蛋白酶變異體
JPH08332032A (ja) 動物経口物用酵素剤及びその用途
KR20190025101A (ko) 항영양인자 제어 미생물 마이크로박테리움 하이드로더멀 kbcbs1021 균주 및 이를 이용한 어분 대체용 발효 대두박 제조 방법
KR101190790B1 (ko) 신규 바실러스 속 ｃｊｂ－０１ 균주 및 사료 첨가제

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): CN JP KR US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase