CN104418945A - Preparation method of peptide and application of peptide in preparation of medicine and feed additive - Google Patents
Preparation method of peptide and application of peptide in preparation of medicine and feed additive Download PDFInfo
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- CN104418945A CN104418945A CN201310389379.5A CN201310389379A CN104418945A CN 104418945 A CN104418945 A CN 104418945A CN 201310389379 A CN201310389379 A CN 201310389379A CN 104418945 A CN104418945 A CN 104418945A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
- A23K20/147—Polymeric derivatives, e.g. peptides or proteins
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
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Abstract
The invention relates to a preparation method of peptide and application of peptide in preparation of a medicine and a feed additive, relating to a bio-pharmaceutical technology. The peptide prepared by the invention has an amino acid sequence represented by Seq ID No.1; found from cell tests, the peptide has the obvious effect of inhibiting hepatitis B virus; and, found from animal tests, the peptide also has the effect of completely improving animal health. The invention further provides a scheme for preparing peptide through a gene engineering method; and therefore, peptide products above gram grade can be prepared. The peptide prepared by the invention has the effects of resisting virus, completing improving animal health and the like, so that the peptide can be used as a main component for preparing a therapeutic medicine and a health-care product which not only can be used by people but also can be used by animals; and the peptide prepared by the invention is applied to preparing the therapeutic medicine, the health-care product and the feed additive.
Description
First part: technical field
The present invention relates to field of biological pharmacy, particularly relate to a kind of peptide (Mytilin B) and preparing the application in medicine, healthy products and fodder additives, preparation method.
Second section: background technology
Alexin is that the one produced through induction in organism has bioactive micromolecule polypeptide, natural immunity material, and molecular weight, about 3000 ~ 7000, is made up of 20 ~ 60 amino-acid residues, multiple disulfide linkage.This kind of active polypeptide majority has the features such as strong basicity, thermostability and broad-spectrum antimicrobial.Natural alexin is distributed in from plant, lower animal to the nearly all biological group such as Mammals, is the important component part of biological autoprotcrtive system.
Most alexin can effectively kill gram-negative bacteria and gram-positive bacteria.Concentration is that namely the alexin of 10 ~ 100mg/L has lethal effect to various bacteria in vitro, and the concentration of alexin in neutrophil leucocyte is g/L level, considerably beyond above-mentioned numerical value, this shows that alexin may have stronger fungicidal activity in vivo, and research at present finds that the kill capability of alexin to gram-positive bacteria is obviously better than gram-negative bacteria.In vitro, HBD-2 is 0.46nmol/ml to colibacillary medium lethal dose (LD50), and minimal inhibitory concentration (MIC) is 15 μ g/ml, and is 62 μ g/mi to the MIC of Pseudomonas aeruginosa, streptococcus aureus.Experiment in vitro proves that most of alexinic MIC scope is 0.5-10 μm of ol/L.
Alexin can also kill some tunicle viruses, as HIV, simplexvirus, bubble type Stomatovirus, but to invalid without capsid virus.θ-alexin also has anti-virus and toxinicide effect.Alexin is mainly through being combined with virus capsid protein thus causing viral loss of biological activity, and this special role mechanism also makes microorganism not easily produce resistivity to it.Alexin directly can suppress virus, the suppression degree of virus is depended on to the tightness degree of alexin concentration and intramolecular disulfide bond, its antiviral efficacy is equally by the impact of the factors such as time, pH value, ionic strength and temperature, under neutrality and conditions of low ionic strength, alexin has stronger antiviral activity.
Alexin not only directly can resist pathogenic micro-organism, but also has immunoregulation effect.The effect that alexin is transmitted by cell signal, strengthens non-specific immunity cell, especially the activity of scavenger cell and chemotaxis.Alexin can also promote chemotactic and the propagation of body T cell, enhancing body immunne response ability, regulates specific immunity, strengthens living organism Initiative Defense function.Alexin can start acquired immune system, and by innate immunity and the organic connection of acquired immunity as cell surface receptors such as a kind of effector molecule activating macrophage, DC, tracheal epithelial cells.Now prove that some α-alexins, beta-alexin have chemotactic activity to T cell, monocyte and immature DC, can induced monocyte and epithelial cell generation cytokine.The neutrophil leucocyte alexin of people, mouse, pig, rabbit can be induced mast cell degranulation and discharge histamine.Beta-alexin also by combining with human chemokine receptor 6 (CCR6), thus attracts jejune dendritic cell (DC) and memory T cell (Tm) to inflammation part, activating cells immunity and humoral immunization.In addition, alexin directly can also promote supplementing and gathering of infection site neutrophil leucocyte.
The use havoc of the Antibiotic Additive microecological balance of animal intestinal, drug residue also have impact on quality and the human health of livestock product, and derive from shellfish, mammiferous alexin relative molecular mass is less, thermostability and water-soluble all more wicked, can absorb in enteron aisle.Because alexin belongs to polypeptide moiety, be easily easily degraded by proteases in vivo as amino acid, general noresidue in vivo after feed intake.Utilize gene engineering method production environment-friendly type alexin fodder additives or added by daily ration, all can effectively improve immunity, prevent disease and growth promoting effects, as reduced stillborn foetus, mummy and deformity; Improve birth weight and piglet birth uniformity coefficient; Raising weaned piglet is heavy, number of weaned, reduces nursery-age pig sickness rate; Reduce milking sow sickness rate, reduce mother, piglet is various stress; Improve sow subhealth state and breeding difficulty, extend sow working life; Improve child care pig surviving rate, reduce sickness rate; Improve child care daily gain in pigs, reduce feedstuff-meat ratio; Improve immunosuppression, improve immunne response ability, antibody horizontal is more neat.
Be derived from the alexin Mytilin B aminoacid sequence of Mediterranean Sea mussel as shown in Seq ID No.3, it is made up of 34 amino-acid residues, 4 disulfide linkage, composite screw and beta structures.Mytilin B alexin has broad spectrum of activity, can suppress Gram-positive and negative bacterium, has antiviral activity simultaneously.Mytilin B alexin utilize N hold with positive charge and the intensive negative electrostatic charge of the cell membrane phospholipid bilayer of cell of prokaryotic cell prokaryocyte attracting, the insertion hydrophobic region that C end is flexible, under amphipathic alpha-helix effect, prokaryotic cell prokaryocyte film is formed the ionic channel of 4nm, the formation of ionic channel changes the osmotic pressure inside and outside cytolemma, in cytolemma, material exosmoses in a large number, causes bacterial cell death, plays germicidal action.The alexinic antiviral-mechanism of Mytilin B is the capsid specific binding with virus, thus suppresses copying of virus.
French scientist has done a large amount of work in the alexin Mytilin research of mussel, natural mussel defence have Mytilin A (Seq ID No.3), Mytilin B (Seq ID No.1), Mytilin C (Seq ID No.4), Mytilin D (Seq ID No.5) and Mytilin G (Seq ID No.6) etc.Because alexin Mytilin molecule is little, there is certain difficulty in separating-purifying, and natural resource are limited, is difficult to industrialization; Because alexinic disulfide linkage and polypeptide space structure determine its basic characteristic, chemosynthesis can not obtain the product with excellent activity, and research shows in many ways, one of biologically active prod percentage only having natural product of chemosynthesis or lower; Only have genetically engineered approach could obtain the alexin with excellent activity, because high density Mytilin alexin polypeptide can kill most host cell, simultaneously in secreting, expressing process, alexin easily degrade by the enzyme system of Host Strains, thus produce internecine situation and cannot really obtain enough expression products.So, also there is no the engineering bacterium expression industrialization production technology of Mytilin alexin polypeptide at present, do not see and accurately express and obtain this enough polypeptide with the report be for experiment, not seen in its application report in treating hepatitis B, immune drug, fodder additives etc. yet.
Part III: summary of the invention
The present invention, according to the blank in above-mentioned field and demand, provides a kind of and is derived from the alexinic biotechnology preparation method of Mediterranean Sea mussel and is preparing the application in medicine, healthy products and fodder additives.
A kind of peptide, is characterized in that having the aminoacid sequence shown in Seq ID No.1.
To encode the gene of above-mentioned peptide.
The nucleotide sequence of described gene is as shown in Seq ID No.2.
Expression vector containing said gene.
The skeleton carrier of described expression vector is pKLAC1.
The preparation method of above-mentioned peptide, is characterized in that above-mentioned expression vector to be transferred in yeast cell, fermentation culture.
Described yeast cell is protease-deficient K.lactis YCT389, YCT390, YCT569 and YCT598.
Above-mentioned peptide is preparing the application in medicine, healthy products and fodder additives, preparation method.
Described medicine and product are oral preparations.
The present invention, according to the alexin Mytilin B being derived from Mediterranean Sea mussel, is made up of 34 amino acid, and molecular weight is 3981.7Da, iso-electric point 9.58, hydrophobic ratio 41%, net charge+9.Due to most polypeptide, comprise Mytilin B can be degraded when yeast secreted expression heterologous protein, its major cause causes due to aspartyl protease effect in Secretory Pathway, and in many cases, only aspartyl protease is degraded with just can causing most of being destroyed property of albumen.K.lactis YCT389, YCT390, YCT569 and YCT598 bacterial strain are specific secretion approach aspartyl protease gene marker-free deletion type, we have selected its Host Strains as Mytilin B, obtain economic and stable expression, realize industrialized manufacture.Our whole process of fermenting process have employed the degraded situation of the polypeptide that LC-MS monitoring secreting, expressing goes out.
The invention provides the preparation method of Mytilin B peptide, the present invention is transferred in yeast expression system by building the expression vector that obtains, makes it express Mytilin B peptide, obtains Mytilin B peptide sterling by follow-up separation and purification drying step.The competent yeast cells K.lactis YCT389 that the present invention preferably adopts, protease deficiency type, specific secretion approach aspartyl protease gene marker-free deletion type, does not have genetic marker, is applicable to the conversion of the expression vector based on linearizing pKLAC or pKLMF.Use this expression system, can guarantee that the Mytilin B peptide of secreting, expressing can not be degraded, possesses activity simultaneously, this expression system also industrialization can express Mytilin A (Seq ID No.3), Mytilin C (Seq ID No.4), MytilinD (Seq ID No.5) and Mytilin G (Seq ID No.6), makes it can be used for medicine manufacture, healthy products and fodder additives.
Test proves, the productive rate of the Mytilin B peptide that the present invention adopts gene engineering method to prepare was more than 0.2% (referring to the ratio of expression product compared with the substratum quality of giving money as a gift that purifying obtains).
The aminoacid sequence of the present invention according to above-mentioned Seq ID No.1, devise the gene order of this aminoacid sequence of coding, utilize the degenerate of codon, have adjusted nucleotide sequence to make it meet yeast expression system to obtain the nucleotide sequence shown in SeqID No.2 to the Preference of codon, be conducive to stability and high efficiency and express alexin.
Present invention also offers the expression vector containing said gene, the skeleton carrier adopted is that the structure of pKLAC1 or pKLAC2, pKLAC2 is shown in Figure of description 1, and by pKLAC1 or pKLAC2, recombinant protein wherein can at cell inner expression, also can secreting, expressing.Be conducive to the alexin obtaining high yield and purity.
Mytilin B peptide provided by the invention as the main component of medicine, healthcare product and fodder additives, can be prepared into medicine and the product of various formulation.As hepatitis B medicine, antiviral, immunostimulant product, sleep improvement product, fodder additives, functional food, veterinary drug, growth stimulant etc.Drug testing in vitro shows, Mytilin B peptide has obvious restraining effect to hepatitis B virus activity, sees embodiment 2.Animal experiment shows, it improves the effect of animal health status in addition comprehensively, sees embodiment 3.The present invention is preferentially prepared into oral preparations.
Part IV: accompanying drawing explanation
Fig. 1. skeleton carrier pKLAC1 structure iron
New England Biolabs, Inc provides
Fig. 2. the plasmid sequencer address of Mytilin B peptide prepared by the present invention
Enzyme is cut the clone of the correct recombinant plasmid pMD19-T of qualification, clone is entirely true in sequencing result display.
Part V: embodiment
The preparation of embodiment 1.Mytilin B peptide
Step 1. foreign DNA synthesizes
Utilize DNA chemosynthesis instrument to prepare foreign gene, obtain the exogenous dna fragment of coding Mytilin B peptide, as shown in SequenceID No.2.
Checking: cut by enzyme and identify that correct clone delivers to promise match genome research center, Beijing and carries out sequencing, sequencer address is shown in (Fig. 2).
The structure of step 2. expression vector
The pKLAC1 (being purchased from New England Biolabs, Inc) that expression vector provides for K.lactis protein expression test kit on pKLAC1 (9097bp), Lac4 promotor (P
lAC4-PB1) 5 ' and 3 ' to hold separate by the replication origin of the gene of encoding beta-lactamase (ApR) and pMB1.K.lactis α-binding factor secretion homing sequence (α-MF), multiple clone site (MCS) and Lac4 transcription terminator (TT) is positioned at 3 ' P
lAC4-PB1downstream; The P of yeast
aDH2the expression of regulation and control acetamidase marker gene (amdS).
Cut carrier pKLAC1 with Sac II or Bst XI enzyme and make its linearizing, exogenous dna fragment Sequence ID No.2 is inserted into the Lac4 promotor site of yeast K.lactis genome self, obtain recombinant vectors.In this recombinant vectors, goal gene is cloned into K.lactis α binding factor (the α Mating Factor on pKLAC1, α-MF) downstream, see Fig. 1, in the nucleus of yeast K.lactis, DNA and the Yeast genome of coding a-MF structural domain and foreign recombinant proteins are integrated, and the α-MF of coding is sheared to make target protein matter secreting, expressing the most at last on golgi body.
P
lAC4-PB1promoter regulation is expressed: once fused protein starts to express, the signal peptide be positioned on α-MF will instruct fused protein to be transported on endoplasmic reticulum (ER) film, is excised by signal peptide at this signal peptidase (SP).Fused protein is transported to golgi body by secretory vesicle (annular), and α-MF structural domain excises at this by Kex proteolytic enzyme, the target protein matter that release is ripe.Subsequently, target protein matter is transported to plasma membrane (PM) by secretory vesicle, is secreted into born of the same parents from plasma membrane.
Step 2. transforms
Use K.lactis YCT389, YCT390, YCT569 and YCT598 (being purchased from New England Biolabs, Inc) competent cell.
Step 1 is built the recombinant vectors obtained and is placed in competent cell solution, make recombinant plasmid be adsorbed in the surface of competent cell, ice bath for some time, cytolemma is in contraction schedule.Then rapid competent cell is placed in 42 DEG C, makes cell membrane channel in thermal environment open, recombinant plasmid by the outer high concentration region of film to film internal diffusion, after 45 second time, put back in ice bath by competent cell again, membrane channel is closed, and recombinant plasmid proceeds to yeast.
Yeast transformant ethanamide screens, and is applied on the basic carbon source of yeast (YCB) substratum containing 5mM ethanamide by cell transformation mixture (containing being transformed by pKLAC1 in a large number or unconverted cell).YCB substratum contains all nutritive substances needed for K.lactis Growth of Cells and carbon source, but lacks nitrogenous source.Only have when ethanamide is degraded to ammonia by acetamidase (the amdS genetic expression by pKLAC1), could be utilized as nitrogenous source.Therefore, the cell of conversion is only had just can to grow up to bacterium colony.
Checking:
Picking colony, with shake-flask culture, substratum is the YPGal substratum (1% yeast extract, 2% peptone, 2% semi-lactosi) containing 38mM ammonium sulfate, 28 DEG C ~ 30 DEG C, incubation time 70 hours.Centrifugal 3000rpm, gets supernatant liquor, agarose gel electrophoresis analysis.
The cultivation of step 3. transformant
Containing YPGal substratum (2% yeast extract of 38mM ammonium sulfate, 4% peptone, 6% semi-lactosi), standard fermentor deep ventilation is cultivated, and 120 DEG C of wet-hot steam sterilizings 30 minutes, are cooled to 28 DEG C, inoculation, 28 DEG C ~ 30 DEG C, in fermenting process, stream mends semi-lactosi 6% again, incubation time 70 hours.
The separation and purification of step 4Mytilin B peptide
Fermented liquid adopts ceramic membrane separation to fall thalline; Then protein content is concentrated to about 5% with nanofiltration; Be heated to 80 degree, keep 20 minutes, be cooled to 20 degree, centrifugation impurity elimination, rotating speed 10000rpm, 30min; Add 10 times amount PH5 hydrochloric acid soln dilutions, make protein content about 0.5%, cross DEAE and G50 gel column; Nanofiltration proteins concentrate content is about 5%, and freeze-drying, product purity is 95%, and purified product is compared with the substratum quality of giving money as a gift, and the productive rate of Mytilin B peptide is about 0.2%
Productive rate={ (fermented liquid expression product total mass 1 × nanofiltration concentration yield × impurity elimination yield × gel separation yield × concentration yield × freeze-drying yield)/give money as a gift substratum total mass } × 100%.
Step 5. is mixed with reserve liquid
The purified of upper step freeze-drying, uses 0.1N dissolving with hydrochloric acid, makes the Mytilin B peptide of 20mg/ml, saves backup.
Embodiment 2.Mytilin B peptide suppresses hepatitis B replication activity test
Step 1. drug treating
Get one bottle, HepG2.2.15 cell, be prepared into single cell suspension with after 0.25% trysinization, after counting, adjustment cell concn is 5 × 10
4cell/ml, 1ml/ hole adds in 24 well culture plates, puts 37 DEG C, 5%CO
2overnight incubation in cell culture incubator.Supernatant is abandoned in suction in second day, adds the DMEM substratum containing different pharmaceutical concentration (50,25,12.5,6.25,3.13 and 0 μMs), often organizes 4 multiple holes.Change the cell culture fluid of same drug level after each effect 4d, and in the 4th and 8 days, collect each hole supernatant in 1.5ml centrifuge tube ,-20 DEG C frozen for subsequent use.
The detection of HBsAg and E antigen in step 2. cells and supernatant
Adopt ELISA method to detect HBsAg and HBeAg, concrete operations are carried out according to the operation instruction of test kit.After color reaction terminates, detect each hole OD value by microplate reader at 450nm place.
Step 3. data processing
Data are expressed as means ± SD.
IC (Inhibition rate, inhibiting rate)=(1-OD
drug treating group/ OD
negative control group) × 100%
Step 4. experimental result, to the restraining effect of hepatitis B replication
HepG2.2.15 cell, after 24 orifice plate overnight incubation, adds the medicine of different weaker concn, sets cell controls group and positive controls simultaneously.After cultivation 4 and 8d, difference collecting cell supernatant, adopts ELISA method to detect the expression of HBsAg and HBeAg.
Tested material Mytilin B peptide and positive drug 3TC make the HBsAg secretion level decline in various degree in nutrient solution, have the restraining effect of obvious dosage correlation in the concentration range of 3.13 ~ 50 μ g.The OD value of blank group compares with the same period, and each concentration group of Mytilin B peptide obviously declines (p<0.05 or 0.01) (the results are shown in Table 1) at 4d and 8d cell conditioned medium HBsAg content.Mytilin B peptide is approximately 3.0 ~ 12 μ g/ml to the half-inhibition concentration IC50 that cell conditioned medium HBsAg secretes.
Table 1Mytilin B peptide is to the restraining effect of HepG2.2.15 emiocytosis HBsAg
Compare with blank group, * p<0.05, * * p<0.01; Compare with the positive controls under same concentration, #p<0.05, ##p<0.01.
Compared with blank group, Mytilin B supernatant HBeAg content when 4d slightly declines, and comparatively obviously (the results are shown in Table 2), the restraining effect of Mytilin B presents certain dosage correlation to restraining effect.(the results are shown in Table 2).
Table 2Mytilin B peptide is to the restraining effect of HepG2.2.15 emiocytosis HBeAg
Compare with blank group, * p<0.05, * * p<0.01; Compare with the positive controls under same concentration, #p<0.05, ##p<0.01.
Tested material Mytilin B peptide and positive drug 3TC have inhibition in various degree to the secretion level of HBsAg and HBeAg in nutrient solution, and Mytilin B peptide presents the relevant restraining effect of certain dosage in the concentration range of 3.13 ~ 50 μ g.
30 days feeding studys of embodiment 3.Mytilin B peptide piglet
The selection of step 1. piglet: the piglet choosing the child care stage, test group and each 30 of control group
Step 2. is tested by Mytilin B peptide quality and standard: non-purifying Mytilin B peptide crude product, net content 1 g/kg, containing dead thalline and indigested substratum.
The feeding conventional baby pig feedstuff of step 3. feeding patterns test group, adds the Mytilin B peptide crude product of 0.5%; The feeding conventional baby pig feedstuff of control group.
Step 4. observation index: the food consumption of piglet, piglet starting weight, the diarrhoea situation of piglet, surviving rate, weightening finish, hair color, the length of one's sleep, feed record.
Step 5. test-results: add the test group piglet that Mytilin B peptide crude product is brought up, hair color, body weight, to search for food and body weight increase is obviously better than control group.In table 3
30 days feeding studys of table 3Mytilin B peptide piglet
Claims (9)
1. a mussel peptide, is characterized in that having the aminoacid sequence shown in Seq ID No.1.
2. the gene of coding peptide according to claim 1.
3. gene according to claim 2, its nucleotide sequence is as shown in Seq ID No.2.
4. the expression vector containing gene according to claim 3.
5. expression vector according to claim 4, its skeleton carrier is pKLAC1 or pKLAC2.
6. the aminoacid sequence polypeptide as shown in Seq ID No.1, Seq IDNo.3, Seq ID No.4, Seq ID No.5, Seq ID No.6 adopts yeast K.lactis YCT389 as described in this patent, YCT390, YCT569 and YCT598 preparation, and K.1actis YCT284, YCT799, yeast saccharomyces cerevisiae, bread yeast, Kluyveromyces lactis and the cereuisiae fermentum preparation of similar yeast.
7. the preparation method of aminoacid sequence polypeptide as shown in Seq ID No.1, Seq ID No.3, Seq ID No.4, Seq ID No.5, Seq ID No.6, is characterized in that being that the expression vector of pKLAC1 or pKLAC2 is transferred in yeast cell according to claim 6 and carries out fermentation culture by skeleton carrier.
8. the aminoacid sequence polypeptide as shown in Seq ID No.1, Seq ID No.3, Seq ID No.4, Seq ID No.5, Seq ID No.6 is preparing the application in medicine, healthy products and fodder additives, as hepatitis B medicine, antiviral, immunostimulant product, sleep improvement product, functional food, fodder additives, veterinary drug, growth stimulant etc.
9. application according to claim 8, described medicine and product refer to oral pharmaceutical, injectable drug, solid and liquid product.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104761625A (en) * | 2015-04-24 | 2015-07-08 | 保罗生物园科技股份有限公司 | Application of defensin mNP in promoting weight gain of chickens |
| CN107279460A (en) * | 2017-06-27 | 2017-10-24 | 兰溪市酉泽饲料技术服务有限公司 | Penaeus Vannmei fermented feed |
| WO2019011286A1 (en) * | 2017-07-14 | 2019-01-17 | Jiangyin Usun Pharmaceutical Co., Ltd. | Antiviral use of mussel adhesive proteins |
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| CN1724566A (en) * | 2004-07-21 | 2006-01-25 | 深圳市翰宇生物工程有限公司 | Antibiotic, antiendotoxin allosteric peptide molecule and synthetic method thereof |
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- 2013-09-02 CN CN201310389379.5A patent/CN104418945A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1724566A (en) * | 2004-07-21 | 2006-01-25 | 深圳市翰宇生物工程有限公司 | Antibiotic, antiendotoxin allosteric peptide molecule and synthetic method thereof |
Non-Patent Citations (1)
| Title |
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| CHARLET,M.ET AL.: ""RecName: Full=Mytilin-B"", 《GENPEPT：P81613.1》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104761625A (en) * | 2015-04-24 | 2015-07-08 | 保罗生物园科技股份有限公司 | Application of defensin mNP in promoting weight gain of chickens |
| CN104761625B (en) * | 2015-04-24 | 2019-05-28 | 保罗生物园科技股份有限公司 | Alexin mNP-1 is promoting the application in chicken weight gain |
| CN107279460A (en) * | 2017-06-27 | 2017-10-24 | 兰溪市酉泽饲料技术服务有限公司 | Penaeus Vannmei fermented feed |
| WO2019011286A1 (en) * | 2017-07-14 | 2019-01-17 | Jiangyin Usun Pharmaceutical Co., Ltd. | Antiviral use of mussel adhesive proteins |
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Application publication date: 20150318 |