CN107996826A - A kind of feed addictive containing alexin and preparation method thereof - Google Patents
A kind of feed addictive containing alexin and preparation method thereof Download PDFInfo
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- CN107996826A CN107996826A CN201711022881.7A CN201711022881A CN107996826A CN 107996826 A CN107996826 A CN 107996826A CN 201711022881 A CN201711022881 A CN 201711022881A CN 107996826 A CN107996826 A CN 107996826A
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- alexin
- preparation
- feed addictive
- arg
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/20—Animal feeding-stuffs from material of animal origin
-
- 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
- A23K20/142—Amino acids; Derivatives thereof
-
- 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
- A23K20/163—Sugars; Polysaccharides
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- Life Sciences & Earth Sciences (AREA)
- Polymers & Plastics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Animal Husbandry (AREA)
- Food Science & Technology (AREA)
- Zoology (AREA)
- Biomedical Technology (AREA)
- Physiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention belongs to field of animal feed, and in particular to a kind of preparation method of the feed addictive containing alexin.Feed addictive of the present invention containing alexin, raw material include D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid alexins.The preparation method of the feed addictive containing alexin; step includes the Yeast gene engineering bacterial strain that fermentation carries D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid phylaxin genes; supernatant is taken to carry out ultrafiltration, ultrafiltrate adds protective agent, wall material and carrier, is spray-dried successively.The additive has height antibiotic property using alexin as main component, which has antimicrobial agent extremely strong antibacterial, biocidal efficacies.
Description
Technical field
The invention belongs to field of animal feed, and in particular to a kind of to contain the feed addictive of alexin and its preparation side
Method.
Background technology
In recent years, antibiotic largely uses in livestock products, it, which is remained, may make animal pathogen produce drug resistance, drug resistance
Gene can by converting, transduceing, engaging, the mode such as transposition ask propagation in bacterium, it is also possible to pass through the approach such as food chain and spread
Drug resistant gene, makes the drug resistant gene of bacterium be transmitted mutually between bacterium in bacterium and the ecosystem in bacterium, fauna in crowd,
Thus cause the drug resistance of pathogenic bacteria to increase and cause the Endodontic failure of human and animal's infectious diseases, the health of the mankind is produced
Raw harm.
The content of the invention
The present invention provides a kind of using alexin as natural feed addictive of major ingredient and preparation method thereof, the additive with
Alexin is main component, has height antibiotic property, which has antimicrobial agent extremely strong antibacterial, biocidal efficacies.
Feed addictive of the present invention containing alexin, raw material are defendd including D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-types amino acid
Element.
The preparation method of feed addictive of the present invention containing alexin, step include fermentation and carry D type amino
The Yeast gene engineering bacterial strain of acid substitution L-type amino acid phylaxin gene, takes supernatant to carry out ultrafiltration, ultrafiltrate adds guarantor successively
Agent, wall material and carrier are protected, is spray-dried.
Preferably, ultrafiltration molecular cut off is the ultrafiltrate of 2KDa~8KDa.
Preferably, the protective agent includes：
Preferably, the wall material is sodium alginate, and additive amount is 5~10wt% of additive gross weight.
Preferably, the carrier is one or both of cornstarch, maltodextrin, wheat bran, and additive amount is addition
The 20-25wt% of agent gross weight.
Preferably, the condition of spray drying is：160~180 DEG C, 10~30ml/min of charging rate of inlet air temperature, outlet air
65~85 DEG C of temperature.
Compared with prior art, the invention has the advantages that：
The additive has height antibiotic property, which has antimicrobial agent extremely strong using alexin as main component
Antibacterial, biocidal efficacies.
Embodiment
The present invention is explained in detail with reference to embodiments.
Embodiment 1
1st, the preparation of yeast fermentation broth
1) tablet culture rejuvenation：Pichia pastoris (this kind of D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid phylaxin genes will be carried
The preparation method of yeast has document and discloses) strain is inoculated on YPD plating mediums, and 48h are cultivated in 28 DEG C, make Pichia pastoris
Rejuvenation, and single bacterium colony is formed, picking single bacterium is fallen within inoculation medium, 28 DEG C of culture 48h；
2) preparation of first order seed：The Pichia pastoris strain switching of step 1) culture is equipped with 300mlYPD liquid culture mediums
In 2L shaking flasks, 28 DEG C of shaking table culture 48h, to late log phase, obtain first order seed；
3) preparation of secondary seed：First order seed prepared by step 2) is transferred to equipped with 30LYPD seed culture mediums
In 50L seeding tanks, 27 DEG C, rotating speed 600rpm of temperature, cultivates 24h, obtains secondary seed solution.
4) preparation of Pichia pastoris zymotic fluid：Secondary seed solution prepared by step 3) is inoculated into according to 5% inoculum concentration
0.5~0.8m3In the fermentation tank of fermentation medium, 31 DEG C, rotating speed 400rpm of temperature, tank pressure 0.05Mpa, ventilating ratio：1：0.6,
48h is cultivated, collects zymotic fluid.
Fermentation medium during fermentation Pichia pastoris consists of the following compositions：Glucose 18g/L, peptone 20g/L, ox
Meat extract 3g/L, yeast extract 8/L, pH 7.2.
2nd, the preparation of D types 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid alexin
1) prepared by supernatant：The zymotic fluid obtained in 1 is centrifuged into 20min through centrifuge 3000rpm, is removed on microorganism collection
Clearly, 5000rpm centrifuges 15min again, removes microorganism collection supernatant, stand-by.
2) ultrafiltration：2KD ultrafiltration membranes are selected, 4000rpm, centrifuges 40min, collects ultrafiltrate.
3) protectant addition：The component of following ratio is pre-mixed, is added with the 5wt% of ultrafiltrate,
It is sufficiently mixed：
4) wall material is prepared and added：Tap water is warming up to 40 DEG C~50 DEG C, gelatin is slowly added into when opening stirring, makes it
The solution for being made into 10wt% is completely dissolved, the gelatin solution prepared is added by the 1.5wt% of ultrafiltrate and stirs 10min.
5) carrier of 25wt% is added, it is 165 DEG C/85 DEG C to set spray drying tower to pass in and out air temperature, and atomizer rotates frequency
Rate is 35Hz, and charging rate 18ml/min, is spray-dried, and is obtained containing D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid alexins
Feed addictive.
Embodiment 2
1st, the preparation of yeast fermentation broth
1) tablet culture rejuvenation：Pichia pastoris (this kind of D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid phylaxin genes will be carried
The preparation method of yeast has document and discloses) strain is inoculated on YPD plating mediums, and 48h are cultivated in 28 DEG C, make Pichia pastoris
Rejuvenation, and single bacterium colony is formed, picking single bacterium is fallen within inoculation medium, 28 DEG C of culture 48h；
2) preparation of first order seed：The Pichia pastoris strain switching of step 1) culture is equipped with 300mlYPD liquid culture mediums
In 2L shaking flasks, 28 DEG C of shaking table culture 48h, to late log phase, obtain first order seed；
3) preparation of secondary seed：First order seed prepared by step 2) is transferred to equipped with 30LYPD seed culture mediums
In 50L seeding tanks, 27 DEG C, rotating speed 600rpm of temperature, cultivates 24h, obtains secondary seed solution.
4) preparation of Pichia pastoris zymotic fluid：Secondary seed solution prepared by step 3) is inoculated into according to 5% inoculum concentration
0.5~0.8m3In the fermentation tank of fermentation medium, 31 DEG C, rotating speed 400rpm of temperature, tank pressure 0.05Mpa, ventilating ratio：1：0.6,
48h is cultivated, collects zymotic fluid.
Fermentation medium during fermentation Pichia pastoris consists of the following compositions：Glucose 18g/L, peptone 20g/L, ox
Meat extract 3g/L, yeast extract 8/L, pH 7.2.
2nd, the preparation of D types 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid alexin
1) prepared by supernatant：The zymotic fluid obtained in 1 is centrifuged into 20min through centrifuge 3000rpm, is removed on microorganism collection
Clearly, 5000rpm centrifuges 15min again, removes microorganism collection supernatant, stand-by.
2) ultrafiltration：8KD ultrafiltration membranes are selected, 4000rpm, centrifuges 40min, collects ultrafiltrate.
3) protectant addition：The component of following ratio is pre-mixed, is added with the 5wt% of ultrafiltrate,
It is sufficiently mixed：
4) wall material is prepared and added：Tap water is warming up to 40 DEG C~50 DEG C, gelatin is slowly added into when opening stirring, makes it
The solution for being made into 10wt% is completely dissolved, the gelatin solution prepared is added by the 1.5wt% of ultrafiltrate and stirs 10min.
5) carrier of 25wt% is added, it is 165 DEG C/85 DEG C to set spray drying tower to pass in and out air temperature, and atomizer rotates frequency
Rate is 35Hz, and charging rate 18ml/min, is spray-dried, and is obtained containing D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid alexins
Feed addictive.
It is added to feed addictive prepared by Examples 1 and 2 in fish meal, significantly improves the feed intake of fry, is promoted
Efficiency of feed utilization, improves the speed of growth and daily gain of fry.
Claims (7)
1. a kind of feed addictive containing alexin, it is characterised in that raw material is prevented including D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid
Imperial element.
2. the preparation method of the feed addictive containing alexin described in a kind of claim 1, it is characterised in that step includes
Fermentation carries the Yeast gene engineering bacterial strain of D type 49-Phe ,82-Ser,115-Arg,144-Met,145-Asn ,161-Arg,169-Met Human Connective tissue growth factor L-type amino acid phylaxin genes, takes supernatant to carry out ultrafiltration,
Ultrafiltrate adds protective agent, wall material and carrier successively, is spray-dried.
3. the preparation method of the feed addictive containing alexin as claimed in claim 1, it is characterised in that ultrafiltration retention point
The ultrafiltrate that son amount is 2KDa~8KDa.
4. the preparation method of the feed addictive containing alexin as claimed in claim 1, it is characterised in that the protective agent
Including：
5. the preparation method of the feed addictive containing alexin as claimed in claim 1, it is characterised in that the wall material is
Sodium alginate, additive amount are 5~10wt% of additive gross weight.
6. the preparation method of the feed addictive containing alexin as claimed in claim 1, it is characterised in that the carrier is
One or both of cornstarch, maltodextrin, wheat bran, additive amount are the 20-25wt% of additive gross weight.
7. the preparation method of the feed addictive containing alexin as claimed in claim 1, it is characterised in that spray drying
Condition is：160~180 DEG C, 10~30ml/min of charging rate of inlet air temperature, 65~85 DEG C of leaving air temp.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711022881.7A CN107996826A (en) | 2017-10-27 | 2017-10-27 | A kind of feed addictive containing alexin and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711022881.7A CN107996826A (en) | 2017-10-27 | 2017-10-27 | A kind of feed addictive containing alexin and preparation method thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107996826A true CN107996826A (en) | 2018-05-08 |
Family
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711022881.7A Pending CN107996826A (en) | 2017-10-27 | 2017-10-27 | A kind of feed addictive containing alexin and preparation method thereof |
Country Status (1)
| Country | Link |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907583A (en) * | 2012-10-27 | 2013-02-06 | 北京大北农科技集团股份有限公司 | Porcine beta defensin-2 spray-dried powder, feed additive, premix and batch |
| CN103703140A (en) * | 2011-03-30 | 2014-04-02 | 德克萨斯大学系统董事会 | Methods and compositions for targeting adipose cells in mammals |
-
2017
- 2017-10-27 CN CN201711022881.7A patent/CN107996826A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103703140A (en) * | 2011-03-30 | 2014-04-02 | 德克萨斯大学系统董事会 | Methods and compositions for targeting adipose cells in mammals |
| CN102907583A (en) * | 2012-10-27 | 2013-02-06 | 北京大北农科技集团股份有限公司 | Porcine beta defensin-2 spray-dried powder, feed additive, premix and batch |
Non-Patent Citations (1)
| Title |
|---|
| 杨星剑等: "抗菌肽的研究进展（上）--基本特性及其活性影响因素", 《农业新技术（今日养猪业）》 * |
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Application publication date: 20180508 |