CN107022537A - A kind of keratinase compound and preparation method and application - Google Patents
A kind of keratinase compound and preparation method and application Download PDFInfo
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- CN107022537A CN107022537A CN201710122600.9A CN201710122600A CN107022537A CN 107022537 A CN107022537 A CN 107022537A CN 201710122600 A CN201710122600 A CN 201710122600A CN 107022537 A CN107022537 A CN 107022537A
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- Prior art keywords
- keratinase
- compound
- amylase
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- 108010059345 keratinase Proteins 0.000 title claims abstract description 58
- 150000001875 compounds Chemical class 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 239000004382 Amylase Substances 0.000 claims abstract description 19
- 108010065511 Amylases Proteins 0.000 claims abstract description 19
- 102000013142 Amylases Human genes 0.000 claims abstract description 19
- 235000019418 amylase Nutrition 0.000 claims abstract description 19
- 238000009472 formulation Methods 0.000 claims abstract description 11
- 239000000203 mixture Substances 0.000 claims abstract description 11
- 241001465754 Metazoa Species 0.000 claims abstract description 9
- 241000186046 Actinomyces Species 0.000 claims abstract description 7
- 244000144972 livestock Species 0.000 claims abstract description 6
- 244000144977 poultry Species 0.000 claims abstract description 5
- 238000007873 sieving Methods 0.000 claims abstract description 4
- 238000000227 grinding Methods 0.000 claims abstract description 3
- 239000007787 solid Substances 0.000 claims description 15
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 10
- 238000011081 inoculation Methods 0.000 claims description 9
- 230000001580 bacterial effect Effects 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 5
- 229910052799 carbon Inorganic materials 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 claims description 5
- 239000002054 inoculum Substances 0.000 claims description 5
- 230000002906 microbiologic effect Effects 0.000 claims description 5
- 229910052757 nitrogen Inorganic materials 0.000 claims description 5
- 241000186361 Actinobacteria <class> Species 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 claims description 2
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 229920002472 Starch Polymers 0.000 claims description 2
- 239000008107 starch Substances 0.000 claims description 2
- 108091005804 Peptidases Proteins 0.000 claims 1
- 102000035195 Peptidases Human genes 0.000 claims 1
- 235000019833 protease Nutrition 0.000 claims 1
- 201000010099 disease Diseases 0.000 abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 9
- 241000187438 Streptomyces fradiae Species 0.000 abstract description 8
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 4
- 230000001151 other effect Effects 0.000 abstract description 3
- 241000193749 Bacillus coagulans Species 0.000 abstract 1
- 229940054340 bacillus coagulans Drugs 0.000 abstract 1
- 241000282898 Sus scrofa Species 0.000 description 10
- 230000003115 biocidal effect Effects 0.000 description 4
- 239000007789 gas Substances 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 102000002568 Multienzyme Complexes Human genes 0.000 description 1
- 108010093369 Multienzyme Complexes Proteins 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 108010019077 beta-Amylase Proteins 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 235000021393 food security Nutrition 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000000505 pernicious effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- GOLXNESZZPUPJE-UHFFFAOYSA-N spiromesifen Chemical compound CC1=CC(C)=CC(C)=C1C(C(O1)=O)=C(OC(=O)CC(C)(C)C)C11CCCC1 GOLXNESZZPUPJE-UHFFFAOYSA-N 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
- C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
- C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
- C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
- C12N9/2405—Glucanases
- C12N9/2408—Glucanases acting on alpha -1,4-glucosidic bonds
- C12N9/2411—Amylases
- C12N9/2425—Beta-amylase (3.2.1.2)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y302/00—Hydrolases acting on glycosyl compounds, i.e. glycosylases (3.2)
- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01002—Beta-amylase (3.2.1.2)
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Enzymes And Modification Thereof (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a kind of bacillus coagulans compound and preparation method and application, described keratinase compound includes 50 ~ 70 parts of parts by weight keratinase, 0 ~ 150 part of amylase 8.The preparation method of described keratinase compound is to weigh keratinase and amylase by formulation ratio, is prepared through low-temperature grinding sieving mixing.Using the application for described keratinase compound in livestock and poultry, aquatic animal feed is prepared.Keratinase in inventive formulation is to purify to obtain from actinomyces, is mainly obtained with the streptomyces fradiae purifying in actinomyces；Inventive formulation can reach promotion growth of animal, the reduction disease of digestive tract incidence of disease, reduction pig house harmful gas concentration and other effects.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of keratinase compound and preparation method and application.
Background technology
With the fast development of animal husbandry, a large amount of of the medicine such as antibiotic use, and feed safety (food security) problem is
As Swine Production person and consumer's focus of attention.World Developed Countries government has assigned in succession to be ordered within a certain time antibiotic usage
And ban.European countries completely forbid nearly ten years adds antibiotic in feed, and China ratifies the antibiosis used in feed
Plain species is also being gradually decreased.Fermented feed can effectively suppress Escherichia coli in digesting and assimilating, salmonella and some causes
Characteristic of disease bacterium and the breeding and growth of pathogen, have certain effect, therefore, countries in the world are right in recent years to the defence of pig disease
Pig fermented feed is especially paid attention to.Existing many pig farms such as Europe state wheat, Holland, Britain are using fermented feed at present, according to statistics,
Fermented feed is being applied on the pig farm that Denmark has 60% so far, and the farm used is also constantly increasing.The U.S. and Canada
To using fermented feed also in popularization energetically.
In recent years, fermented feed has also obtained preferable development in China.It is reachable using animals such as the feeding pigs of fermented feed
To promotion growth of animal, the reduction disease of digestive tract incidence of disease, reduction pig house harmful gas concentration and other effects.Therefore, develop a kind of
Fermented feed raw material is very important.
The content of the invention
The first object of the present invention is to provide a kind of keratinase compound；Second purpose is to provide the keratin
The preparation method of multienzyme complex；3rd purpose is that providing the keratinase compound is preparing livestock and poultry, aquatic animal feed
In application.
The first object of the present invention is achieved in that described keratinase compound includes parts by weight keratinase 50
~70 parts, 0~150 part of amylase 8.
The second object of the present invention is achieved in that weighs keratinase and amylase by formulation ratio, through low temperature powder
Broken sieving mixing is prepared.
The third object of the present invention is achieved in that described keratinase compound is preparing livestock and poultry, aquatic livestock
Application in feed.
Keratinase in inventive formulation is to purify to obtain from actinomyces, mainly with the streptomyces fradiae in actinomyces
Purifying is obtained；It is dense that inventive formulation can reach promotion growth of animal, the reduction disease of digestive tract incidence of disease, reduction pig house pernicious gas
Degree and other effects.
Embodiment
The present invention is further illustrated below, but the present invention is not any limitation as in any way, based on the present invention
Any conversion or replacement that training centre is made, belong to protection scope of the present invention.
Keratinase compound of the present invention includes 50~70 parts of parts by weight keratinase, 0~150 part of amylase 8.
Described keratinase compound includes 55~65 parts of parts by weight keratinase, 90~120 parts of amylase.
Described keratinase compound includes 60 parts of parts by weight keratinase, 100 parts of amylase.
Described keratinase is to purify to obtain from actinomyces.
The method of described purifying comprises the following steps：
1) using the inoculation of conventional microbiological inoculation method：Test tube slant culture actinomyces, using Cha Shi solid mediums；Move
Connect：By the actinomycetes strain transposing of above-mentioned growth into production spore solid medium, 30~35 DEG C, cultivate 2~7 days, to spore production
It is raw；
2) bacterial strain spore in 1) step is added in solid medium by 2~5% inoculum concentrations, controls culture medium carbon: nitrogen
Ratio in (2~4)：In the range of (1~1.5), Phos is added, its content is 5~8 μm of ol/g butts, keeps humidity 40
~60%, 30~35 DEG C of temperature, natural ph, keratinase generation time is 40~80 hours.
Described amylase is beta amylase.
The preparation method of keratinase compound of the present invention, is to weigh keratinase and starch by formulation ratio
Enzyme, is prepared through low-temperature grinding sieving mixing.
The application of keratinase compound of the present invention is that described keratinase compound is preparing livestock and poultry, aquatic products
Application in animal feed.
With specific embodiment, the present invention will be further described below：
Embodiment 1
1) using the inoculation of conventional microbiological inoculation method：Test tube slant culture streptomyces fradiae, using Cha Shi solid cultures
Base；Transposing：By the streptomyces fradiae bacterial strain transposing of above-mentioned growth into production spore solid medium, 35 DEG C are cultivated 7 days, to spore production
It is raw；
2) bacterial strain spore in 1) step is added in solid medium by 5% inoculum concentration, controls culture medium carbon: the ratio of nitrogen
Example is 2：1.5, Phos is added, its content is 8 μm of ol/g butts, keep humidity 50~60%, 30~35 DEG C of temperature is natural
PH value, keratinase generation time is 80 hours.
Embodiment 2
1) using the inoculation of conventional microbiological inoculation method：Test tube slant culture streptomyces fradiae, using Cha Shi solid cultures
Base；Transposing：By the streptomyces fradiae bacterial strain transposing of above-mentioned growth into production spore solid medium, 30 DEG C, cultivate 5 days, to spore
Produce；
2) bacterial strain spore in 1) step is added in solid medium by 2% inoculum concentration, controls culture medium carbon: the ratio of nitrogen
Example is 4：In the range of 1, Phos is added, its content is 5 μm of ol/g butts, keeps humidity 40~50%, temperature 30~35
DEG C, natural pH values, keratinase generation time is 40 hours.
Embodiment 3
1) using the inoculation of conventional microbiological inoculation method：Test tube slant culture streptomyces fradiae, using Cha Shi solid cultures
Base；Transposing：By the streptomyces fradiae bacterial strain transposing of above-mentioned growth into production spore solid medium, 33 DEG C, cultivate 2 days, to spore
Produce；
2) bacterial strain spore in 1) step is added in solid medium by 4% inoculum concentration, controls culture medium carbon: the ratio of nitrogen
Example is 3：In the range of 1.2, Phos is added, its content is 6 μm of ol/g butts, keeps humidity 45~55%, temperature 30~35
DEG C, natural ph, keratinase generation time is 50 hours.
Embodiment 4
The keratinase that embodiment 2 is prepared is weighed by the formulation ratio of keratinase and amylase, by keratinase
100 mesh sieves were crushed below 30 DEG C of temperature be uniformly mixed so as to obtain object with amylase.
Embodiment 5
The keratinase that embodiment 1 is prepared is weighed by the formulation ratio of keratinase and amylase, by keratinase
100 mesh sieves were crushed below 30 DEG C of temperature be uniformly mixed so as to obtain object with amylase.
Embodiment 6
The keratinase that embodiment 3 is prepared is weighed by the formulation ratio of keratinase and amylase, by keratinase
100 mesh sieves were crushed below 30 DEG C of temperature be uniformly mixed so as to obtain object with amylase.
Embodiment 7
Pig feed is made in the keratinase compound for respectively preparing embodiment 4, embodiment 5 and embodiment 6, through feeding
The growth of pig has obtained larger promotion, while basic do not have disease of digestive tract morbidity, pig house harmful gas concentration big after supporting
Width is reduced.
Claims (8)
1. a kind of keratinase compound, it is characterised in that described keratinase compound include parts by weight keratinase 50 ~
70 parts, 0 ~ 150 part of amylase 8.
2. keratinase compound according to claim 1, it is characterised in that described keratinase compound includes weight
Measure 55 ~ 65 parts of part keratinase, 90 ~ 120 parts of amylase.
3. keratinase compound according to claim 1, it is characterised in that described keratinase compound includes weight
Measure 60 parts of part keratinase, 100 parts of amylase.
4. the keratinase compound according to claim 1,2 or 3, it is characterised in that described keratinase is from unwrapping wire
Purifying is obtained in bacterium.
5. frontal eminence proteinase complex according to claim 4, it is characterised in that the method for described purifying includes following step
Suddenly：
1）It is inoculated with using conventional microbiological inoculation method：Test tube slant culture actinomyces, using Cha Shi solid mediums；Transposing：
By the actinomycetes strain transposing of above-mentioned growth into production spore solid medium, 30 ~ 35 DEG C, cultivate 2 ~ 7 days, produced to spore；
2）By 1）Bacterial strain spore is added in solid medium by 2 ~ 5% inoculum concentrations in step, controls culture medium carbon: the ratio of nitrogen
（2 ~ 4）：（1 ~ 1.5）In the range of, Phos is added, its content is 5 ~ 8 μm of ol/g butts, keeps humidity 40 ~ 60%, temperature
30 ~ 35 DEG C of degree, natural ph, keratinase generation time is 40 ~ 80 hours.
6. the keratinase compound according to claim 1,2 or 3, it is characterised in that described amylase is β-starch
Enzyme.
7. the preparation method of any described keratinase compound of a kind of claim 1 ~ 7, it is characterised in that by formulation ratio
Keratinase and amylase are weighed, is prepared through low-temperature grinding sieving mixing.
8. a kind of application of any described keratinase compound of claim 1 ~ 7, it is characterised in that described keratinase
Application of the compound in livestock and poultry, aquatic animal feed is prepared.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710122600.9A CN107022537A (en) | 2017-03-03 | 2017-03-03 | A kind of keratinase compound and preparation method and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710122600.9A CN107022537A (en) | 2017-03-03 | 2017-03-03 | A kind of keratinase compound and preparation method and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107022537A true CN107022537A (en) | 2017-08-08 |
Family
ID=59525423
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710122600.9A Pending CN107022537A (en) | 2017-03-03 | 2017-03-03 | A kind of keratinase compound and preparation method and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107022537A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1417328A (en) * | 2001-11-09 | 2003-05-14 | 中国科学院沈阳应用生态研究所 | Solid fermentation prepn process and application of keratinase |
| CN102669443A (en) * | 2002-08-09 | 2012-09-19 | 北卡罗莱纳州立大学 | Methods and compositions for improving growth of meat-type poultry |
| CN103533843A (en) * | 2011-02-18 | 2014-01-22 | 杜邦营养生物科学有限公司 | Feed additive composition |
-
2017
- 2017-03-03 CN CN201710122600.9A patent/CN107022537A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1417328A (en) * | 2001-11-09 | 2003-05-14 | 中国科学院沈阳应用生态研究所 | Solid fermentation prepn process and application of keratinase |
| CN102669443A (en) * | 2002-08-09 | 2012-09-19 | 北卡罗莱纳州立大学 | Methods and compositions for improving growth of meat-type poultry |
| CN103533843A (en) * | 2011-02-18 | 2014-01-22 | 杜邦营养生物科学有限公司 | Feed additive composition |
Non-Patent Citations (3)
| Title |
|---|
| HAIYING WANG ET AL.: "Effects of dietary supplementation of keratinase on growth performance, nitrogen retention and intestinal morphology of broiler chickens fed diets with soybean and cottonseed meals", 《ANIMAL FEED SCIENCE AND TECHNOLOGY》 * |
| 李明珠 等: "角蛋白酶的研究现状及发展前景", 《安徽农业科学》 * |
| 李虓 等: "多酶生物饲料的研究进展", 《粮食与饲料工业》 * |
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Application publication date: 20170808 |
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