CN1286525C - New type pellicle of ocean lysozyme and preparation method - Google Patents
New type pellicle of ocean lysozyme and preparation method Download PDFInfo
- Publication number
- CN1286525C CN1286525C CN 200510000033 CN200510000033A CN1286525C CN 1286525 C CN1286525 C CN 1286525C CN 200510000033 CN200510000033 CN 200510000033 CN 200510000033 A CN200510000033 A CN 200510000033A CN 1286525 C CN1286525 C CN 1286525C
- Authority
- CN
- China
- Prior art keywords
- lysozyme
- ocean
- ocean lysozyme
- filmogen
- pellicle
- Prior art date
- Legal status (The legal status 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 status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The present invention relates to a novel ocean lysozyme film agent, which can be obtained by preparing and film coating ocean lysozyme, film forming material, a plasticizer and water. The film agent has strong killing action to gram-positive and gram-negative bacteria in clinic. The antimicrobial spectrum is wide, and the curing degree to rat vaginas of mixed infection by bacteria is 92.9%. The present invention has no poison and side effect, and no stimulation action. The present invention is a new generation of novel ocean lysozyme film agent.
Description
Technical field
The present invention relates to the medical drugs preparation, particularly relate to pellicle of ocean lysozyme that produces by Marine microorganism bacillus cereus S-12 and preparation method thereof.
Background technology
Along with development of biology, seek the focus that the enzyme with brand-new character has become international enzyme preparation research in recent years.Marine organisms are owing to be grown in this unique environment of ocean, particularly live in Marine microorganism and microalgae under the extreme environment, can produce abundant extreme enzyme, bring up Marine microorganism and be different from all polyspecifics of land microorganism, compare with the enzyme that land Institute of Micro-biology produces, the ocean enzyme has more unique zymologic property, has therefore caused the very big concern of domestic and international research worker.
At present, the comparatively thorough microbial source lysozyme of research mainly contains the M1 enzyme (Lichenstein H S 1990) of ChL (Lyne J E 1990) the .Streptomyces globisporus generation from Chalaropsis, the Cellosyl (Astid R 2001) that S.coelicolor produces etc.In this bacteriolyze system, contain 3 kinds of hydrolytic enzyme at least with bacteriolysis.When the hydrolytic bacteria cell wall, these different lysozyme act on the different chemical site of whole cell peptidoglycan respectively, finish " molten wall " this biological process jointly.It can cut off the contact of β-1,4 glycosidic bond between the N-acetylglucosamine and-acetylmuramic acid in the bacteria cell wall Peptidoglycan, destroys the Peptidoglycan support, and antibacterial cell spalling under the effect that internal penetration is pressed is opened, and causes the antibacterial cracking and killing bacteria.The acellular wall construction of humans and animals cell does not also have Peptidoglycan, so lysozyme is to the human body cell free of toxic effects.At present, people mainly produce lysozyme from Ovum Gallus domesticus album, yet lysozyme of chicken can cause allergy and can't be used for the mankind as foreign protein.Lysozyme removes directly cracking antibacterial, and the inductive bacterial endotoxin of ceftazidime (LPS) is discharged the activity that inhibitory action is arranged and kill fungus, causes that people greatly pay close attention to.The open CN1302634A of Chinese patent openly uses the treatment by Chinese herbs inflammation of uterus, but drug action is slow, so that state of an illness protracted course of disease or outbreak repeatedly, and clinical effectiveness is bad.The CN1488400A report is made biological preparation with staphylococcus lysozyme lysozyme of chicken and bacteriostatic peptide and is prevented and treat cow endometritis.
Summary of the invention
The objective of the invention is to overcome prior art and exist not enoughly, be engaged in the developmental research of Marine microorganism for a long time through the inventor, exploitation provides a kind of ocean lysozyme of Marine microorganism bacillus cereus S-12 high yield to prepare pellicle of ocean lysozyme and preparation method thereof.
New type pellicle of ocean lysozyme provided by the invention by following component and weight portion through preparation, filming forms:
Ocean lysozyme 3-36
Filmogen 10-55
Plasticizer 16.40-72
Water 1400-1900.
New type pellicle of ocean lysozyme provided by the invention preferably by following component and weight portion through preparation, filming forms
Ocean lysozyme 10-30
Filmogen 20-40
Plasticizer 25.2-63
Water 1500-1800.
New type pellicle of ocean lysozyme provided by the invention more preferably by following component and weight portion through preparation, filming forms
Ocean lysozyme 20.00
Filmogen 33.40
Plasticizer 54.7
Water 1670.
The preparation method of new type pellicle of ocean lysozyme provided by the invention comprises that the following step poly-:
(1) preparation
Above-mentioned formula ratio filmogen is disperseed with boiling water, be stirred to room temperature, become transparent colloidal solution.
Get above-mentioned formula ratio ocean lysozyme, add in the filmogen colloidal solution, add the plasticizer of above-mentioned formula ratio again, add water to above-mentioned prescription Total Water and stir, leave standstill bubble collapse and get colloidal solution with suitable quantity of water dissolving back.
(2) film
In new type pellicle of ocean lysozyme provided by the invention and preparation method thereof, described ocean lysozyme (is deposited in typical case's culture collection center, Chinese Wuhan from a kind of bacillus cereus S-12 strain that culture of isolated obtains high-yield marine microorganism lysozyme (title lysozyme) from the sea mud in marine site, the East Sea, preserving number is CCTCC NO:M204089) ocean lysozyme that produces, this lysozyme physicochemical property is:
The molecular weight of lysozyme is 39KD;
The molecule of lysozyme is made up of 354 aminoacid;
The isoelectric point, IP of lysozyme is pI＞9;
The lysozyme optimum pH is 8;
The optimum temperature of lysozyme is 35 ℃;
Metal ion is to the influence of enzyme activity:
Most metal ion is lived to enzyme does not influence Co
2+, Cu
2+, Ag
+To enzyme slightly activation alive, Mg
2+, Zn
2+, Ca
2+Inhibitory action is arranged;
The pH stability and the heat stability of enzyme: enzyme is good stability between pH4.0-10.0, and enzyme is stable preferably in the time of 45 ℃;
Lysozyme reaches 3279.5u/mg than living behind the purification.
Lysozyme is consumption (weight portion) 3-36 in film, is preferably 20-30, and more preferably 20.00.
Described filmogen is natural or synthetic macromolecular compound, for example starch, dextrin, gelatin, agar, chitin or cellulose and derivant such as hydroxypropyl emthylcellulose etc., be preferably hydroxypropyl emthylcellulose, its consumption (weight portion) is generally 10-55, be preferably 20-40, more preferably 33.40.
Described plasticizer is glycerol or sorbitol etc., is preferably 54.7.
In the new type pellicle of ocean lysozyme provided by the invention, be that (every lattice are 30 centimetres in unit with lattice
2), every lattice contain the 85-115% that ocean lysozyme should be the expression amount, and this membrane is a light brown, can swollen filming in water.Get membrane one lattice, add water 2ml, jolting makes dissolving, adds 4 of 20 of 10% sodium hydroxide solutions and 10% copper-baths, displaing amaranth behind the mixing.
Get membrane one lattice, press N2 method (" Chinese pharmacopoeia version in 2000, two appendix VIID second methods) and measure, nitrogen content should be not less than 1.8mg.
This lysozyme membrane under the listing terms of packing 25 ± 2 ℃, 35 ± 5 ℃, the indoor placement of 60% ± 10%RH 6 months, its outward appearance, content, microbiology detect does not have no significant change, and the confirmation sample is steady quality under these conditions.
Following table 1 is that the result is shown in 25 ± 2 ℃ of surveys.
Table 1
| Detection time | Outward appearance | Content | |
| 0 month | The light brown film, complete bright and clean, there is not obvious bubble | 104.6％ | Up to |
| 1 month | The light brown film, complete bright and clean, there is not obvious bubble | 103.5％ | Up to |
| 2 months | The light brown film, complete bright and clean, there is not obvious bubble | 105.7％ | Up to specification |
| 3 months | The light brown film, complete bright and clean, there is not obvious bubble | 104.3％ | Up to |
| 4 months | The light brown film, complete bright and clean, there is not obvious bubble | 102.2％ | Up to specification |
| 5 months | The light brown film, complete bright and clean, there is not obvious bubble | 102.7％ | Up to specification |
| 6 months | The light brown film, complete bright and clean, there is not obvious bubble | 104.3％ | Up to specification |
One, ocean lysozyme pharmacodynamics test research
The experiment of ocean lysozyme antibacterial activity in vitro
Adopt the plate doubling dilution to measure ocean lysozyme to the clinical common Gram-positive of clinical isolating 160 strains and the vitro antibacterial activity of gram negative bacteria.
Select 8 kind of 160 strain antibacterial altogether for use, be respectively staphylococcus aureus 20 strains, staphylococcus epidermidis 20 strains, colon bacillus 20 strains, Pseudomonas aeruginosa 20 strains, klebsiella pneumoniae 20 strains, proteus vulgaris 20 strains, enterococcus 20 strains, 20 strains of motionless Pseudomonas.Above bacterial strain is the clinical isolates strain that Shandong Qilu Hospital collects in the recent period, all carries out conventional method and identifies.
Staphylococcus aureus ATCC25922, colon bacillus ATCC25923, Pseudomonas aeruginosa ATCC27853 all derive from Nat'l Pharmaceutical ﹠ Biological Products Control Institute, as the Quality Control bacterial strain in the test.
The final concentration of above-mentioned each bacterium experiment is 1 * 10
5CFU/ml.
The result shows that ocean lysozyme has antibacterial preferably or bactericidal effect.To the MIC scope of most of strains all between 0.125-32ug/ml, MIC
50Mostly be 1-16ug/ml greatly, stronger to golden Portugal bacterium, form staph, colon bacillus, Bacillus proteus, enterococcus, klebsiella pneumoniae and the effect of motionless Pseudomonas, effect is taken second place to pseudomonas aeruginosa, MIC
50For 8ug/ml sees Table 2.Ocean lysozyme also has good inhibitory effect to see Table 3 to 3 strain reference cultures.
Ocean lysozyme is comparatively approaching to MBC and the MIC of 8 strain antibacterials, proves that its killing action that act as to experimental strain sees Table 4.
Fig. 1-8 is that ocean lysozyme is to the antibacterial percentage curves of the concentration of 8 kinds of antibacterials.
To sum up the result shows, ocean lysozyme all has stronger killing action to clinical common Gram-positive and gram negative bacteria, demonstrates better antibacterial activity, and antimicrobial spectrum is comparatively extensive.
Table 2 ocean lysozyme is to the MIC measured value of 180 strain antibacterials
| Antibacterial | The strain number | Medicine | MIC scope (ug/ml) | MIC 50 (ug/ml) | MIC 90 (ug/ml) |
| Staphylococcus aureus MRSE ETEC pseudomonas aeruginosa | 20 20 20 20 | Ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene | 0.25-16 0.5-64 0.0625-8 0.03125-4 0.125-32 2-64 0.5-32 2-64 | 4 8 1 1 4 16 8 16 | 16 16 4 4 32 64 16 64 |
| The motionless Pseudomonas enterococcus of proteus pneumonia klebsiella spp | 20 20 20 20 | Ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene ocean lysozyme chlorohexidene | 0.0625-32 0.125-32 0.125-16 0.5-32 0.25-32 0.125-32 0.25-16 0.25-32 | 2 4 2 4 4 4 4 4 | 8 32 16 16 32 16 16 16 |
Table 3 ocean lysozyme is to the MIC measured value of 3 strain reference cultures
| Antibacterial | The strain number | Medicine | MIC(ug/ml) |
| Staphylococcus aureus (ATCC25922) colon bacillus (ATCC25923) Pseudomonas aeruginosa (ATCC27853) | 1 1 1 | Ocean lysozyme hibitane ocean lysozyme hibitane ocean lysozyme hibitane | 0.125 0.25 0.25 0.125 1 0.25 |
Table 4 ocean lysozyme is to the MBC measured value of 8 strain antibacterials
| Antibacterial | Bacterium number | MIC(mg/ml) | MBC(mg/ml) |
| Staphylococcus aureus staphylococcus aureus bacillus pyocyaneus bacillus pyocyaneus escherichia coli | JP-04-08 ATCC25922 LN-04-12 ATCC27853 DC-04-21 | 0.125 0.125 0.0625 0.125 0.25 | 0.5 0.25 0.25 0.5 0.25 |
| Escherichia coli staphylococcus epidermidis proteus vulgaris | ATCC25923 BP-04-18 BX-04-10 | 0.25 0.0625 0.25 | 0.5 0.25 1 |
Two, ocean lysozyme is to the therapeutical effect of rat bacterial vaginitis
By the animal model of rat vagina mixed infection gold Portugal bacterium and colon bacillus, be index with the antibacterial negative conversion rate of vaginal secretions and the healing degree of vaginal mucosa inflammation, observe the therapeutic effect of 3 dosage groups of ocean lysozyme.
Select 2 kinds of vaginal infection common bacteria for use, promptly staphylococcus aureus and colon bacillus are the clinical isolates strain that Shandong Qilu Hospital collects in the recent period.
Animal is the Wistar rat, and is female, body weight: 200-250g, anti-medical Group Co.,Ltd provides by the Shandong, Shandong.
Grouping and treatment
With 50 of the rats of modeling success, be divided into 5 groups respectively at random, promptly the heavy dose of group of ocean lysozyme, middle dosage group, small dose group, hibitane (positive control) group, model group, get 10 normal rats in addition and be blank (normally) matched group.3 dosage groups of ocean lysozyme give ocean lysozyme 10mg/kg, 5mg/kg, 2.5mg/kg respectively, and the hibitane group gives hibitane 5mg/kg, and model group and blank group are washed to warm saline, continuous 7 days.
Put to death rat on the 6th day in drug withdrawal and dissect, get vagina and carry out perusal.And mark by table 3.Score value with model group is an inflammation degree 100%, and administration group and model group relatively can draw the inflammation degree of administration group, and blank group is administration group healing degree with the difference of administration group inflammation degree.
Table 5 rat vagina mucositis sexual stimulus standards of grading
| | Reaction symptom | |
| 0 1 2 | There are not hyperemia, edema, hemorrhage, the level and smooth vaginal mucosa of vaginal mucosa is still level and smooth, there is the mild hyperaemia vaginal mucosa still level and smooth, moderate hyperemia, visible point-like petechia |
| 3 | Vaginal mucosa is unsmooth, severe hyperemia, and Mild edema is organized in visible volume petechia |
Overall score/model group the overall score of inflammation degree=administration group (or positive group)
The inflammation degree of the inflammation degree-administration group of healing degree=model group (or positive group)
(1) ocean lysozyme sees Table 6 to the negative conversion rate of rat vagina bacterial infection.
Table 6 ocean lysozyme is to the negative conversion rate of rat vagina bacterial infection
| Group | Dosage | Number of animals | Negative conversion rate (%) | |
| Staphylococcus aureus | Colon bacillus | |||
| Dosage group ocean lysozyme small dose group positive controls model group in the heavy dose of group of the ocean lysozyme ocean lysozyme | 10mg/kg 5mg/kg 2.5mg/kg 5mg/kg | 10 10 10 10 10 | 80 50 30 60 0 | 90 60 30 80 0 |
(2) ocean lysozyme sees Table 7 to the inflammation score value and the inflammation degree of rat vagina mixed cell infection vagina specimen.
Table 7 ocean lysozyme infects the inflammation score value and the inflammation degree (n=10) of vagina to the rat vagina mixed cell
| Group | Dosage (mg/kg) | The inflammation score value | Inflammation degree (%) | |||
| 1 | 2 | 3 | Amount to | |||
| Dosage group ocean lysozyme small dose group in the heavy dose of group of the ocean lysozyme ocean lysozyme | 10 5 2.5 | 2 (2) 3 (3) 4 (4) | 0 2 (1) 6 (3) | 0 0 0 | 2 5 10 | 7.1 17.9 35.7 |
| Positive controls model group blank group | 5 | 5 (5) 0 0 | 0 4 (2) 0 | 0 24 (8) 0 | 5 28 0 | 17.9 100 0 |
Annotate: (1) is to be the curative effect of number of animals (3) ocean lysozyme to the bacteria mixed infection rat vagina in 100% (2) bracket with model control group inflammation degree, sees Table 8.
Table 8 ocean lysozyme is to the curative effect of bacteria mixed infection rat vagina
| Group | Dosage | Healing degree (%) |
| Dosage group ocean lysozyme small dose group positive controls in the heavy dose of group of the ocean lysozyme ocean lysozyme | 10mg/kg 5mg/kg 2.5mg/kg 5mg/kg | 92.9 82.1 64.3 82.1 |
The result shows that ocean lysozyme 10,5,2.5mg/kg vagina administration are respectively 80%, 50%, 30% to the negative conversion rate that rat vagina gold Portugal bacterium infects, and the negative conversion rate that the rat vagina colon bacillus is infected is respectively 90%, 60%, 30%.Healing degree to the bacteria mixed infection rat vagina is respectively 92.9%, 82.1% and 64.3%, illustrates that ocean lysozyme has bacteria growing inhibiting effect preferably and therapeutical effect in the intravaginal that mixed cell infects, and certain amount-effect dependency is arranged.
The clean film of sea woman is to the rabbit vagina irritant test
The clean film of medicine sea woman, content: 21%, provide lot number by Affiliated Hospital of Qingdao University, Chinese aquatic science institute, the Yellow River Aquatic product institute: 20040512.
Animal: new zealand rabbit
Body weight: 2.50kg ±
Sex: female
Number of animals: 16
The Shandong, Shandong resists medical Group Co.,Ltd.
Route of administration: vagina administration
(1) single-dose
Get 8 rabbit and be divided into 2 groups at random by body weight, the clean film group of sea woman, vehicle group, every group 4, during administration animal is fixed in the high low level of buttocks, the clean film 1.0g/ of extra large woman only is rolled into cylindric about 3.0cm, expand with Nasal dilator and to open vaginal orifice and live the medicine film with ophthalmology tweezers pincers and put into intravaginal 1cm gently, make animal keep a buttocks height low level passive quiet 5 minutes after the administration.The vehicle group animal gives the excipient of equivalent, and method is with medicine film group.Put to death zootomy in 24 hours behind the medicine, observe the medication local mucous membrane and have or not irritant reaction such as hyperemia, edema.
(2) multiple dosing
Get 8 rats and be divided into 2 groups at random by body weight, the clean film group of extra large woman, vehicle group, 4 every group, during administration animal is fixed in the high low level of buttocks, the clean film 1.0g/ of extra large woman only is rolled into cylindric about 3.0cm, put into intravaginal 1cm,
The result shows clean film single of extra large woman and repeatedly rabbit vagina administration, animal generally in order, unusual performance such as no restlessness behind the medicine, local do not have stimulations such as obvious hyperemia, edema through dissecting the vagina perusal, histopathologic examination, vagina epithelium is normal, does not see morphological changes such as hypertrophy, inflammation, necrosis.Rabbit single and continuous use are repeatedly given in the prompting of above-mentioned result of the test, the clean film of extra large woman, and to the animal whole body, the vagina part does not all produce tangible stimulation, illustrates that the clean bolt of extra large woman, the clinical topical application of the clean film of extra large woman are safe.
New type pellicle of ocean lysozyme characteristics provided by the invention are:
1, new type pellicle of ocean lysozyme Chinese medicine lysozyme is from the ocean lysozyme that obtains new bacillus cereus S-12 high yield in the sea mud of the East Sea, and physio-biochemical characteristics uniqueness, particularly pharmacological properties are different from land as lysozyme of chicken.Be a biological agent fully, have vast market prospect.
2, pellicle of ocean lysozyme is stronger to clinical common gram-bacteria, Bacillus proteus, enterococcus, kerekou pneumonia one hundred bacillus and the effect of motionless Pseudomonas, and antimicrobial spectrum is comparatively extensive.
3, ocean lysozyme is 92.9% to the healing degree of bacteria mixed infection rat vagina.
4, this membrane adopts topical, and therapeutical effect is fast, has no side effect and has no stimulation, and is easy to use, and the problem of having no drug resistance is a generation new type pellicle of ocean lysozyme.
Description of drawings
Fig. 1 ocean lysozyme is to the antibacterial percentage rate of golden staphylococcus (20 strain),
The antibacterial percentage rate of Fig. 2 ocean lysozyme his-and-hers watches staphylococcus (20 strain),
Fig. 3 ocean lysozyme is to the antibacterial percentage rate of escherichia coli (20 strain),
Fig. 4 ocean lysozyme is to the antibacterial percentage rate of green amine bacterium (20 strain),
Fig. 5 ocean lysozyme is to the antibacterial percentage rate of proteus vulgaris (20 strain),
Fig. 6 ocean lysozyme is to the antibacterial percentage rate of kerekou pneumonia Bai Shi bacillus bacterium (20 strain),
Fig. 7 ocean lysozyme is to the antibacterial percentage rate of motionless Pseudomonas (20 strain),
Fig. 8 ocean lysozyme is to the antibacterial percentage rate of enterococcus (20 strain),
The specific embodiment
Take by weighing hydroxypropyl emthylcellulose 33.40g, put in the 2000ml beaker, 1000ml is uniformly dispersed with boiling water, stirs, and is placed to room temperature, becomes transparent colloidal solution.
Take by weighing ocean lysozyme 20.00g and put in the beaker, with the suitable quantity of water dissolving, this principal agent solution is transferred in the hydroxypropyl emthylcellulose colloidal solution, with low amounts of water washing beaker, washing liquid is incorporated the hydroxypropyl emthylcellulose colloidal solution into; Add glycerol and water,, stir, make and misciblely evenly leave standstill, film to there not being bubble substantially to water inventory 1670ml.
Wet film was in 38 ~ 40 ℃ of dryings 12 hours, and skinning is done the semi-finished product assay, fixed heavily cutting, packing.
| Example 1 | Example 2 | |
| Outward appearance | The light brown film, the film forming situation is good, and film is smooth, smooth; There is Disabled to stay liquid on the mould | The light brown film, the film forming situation is better, and film is smooth, smooth; There is a little Disabled to stay liquid on the mould |
| Toughness | Greatly | Moderate |
| Swelling time | 25min | 10min |
The step of embodiment 2 is identical with embodiment's 1, is not both glycerol consumption 2 times for embodiment.
Claims (6)
1, a kind of pellicle of ocean lysozyme, by following component and weight portion through preparation, filming forms
Ocean lysozyme 3-36
Filmogen 10-55
Plasticizer 16.40-72
Water 1400-1900;
Described ocean lysozyme is to be the ocean lysozyme of the bacillus cereus S-12 strain product of CCTCC NO:M204089 by preserving number; Described filmogen is starch, dextrin, gelatin, agar, chitin or cellulose and derivant thereof; Described plasticizer is glycerol or sorbitol.
2,, it is characterized in that described each component and weight portion are according to the pellicle of ocean lysozyme of claim 1
Ocean lysozyme 10-30
Filmogen 20-40
Plasticizer 25.2-63
Water 1500-1800;
Described ocean lysozyme is to be the ocean lysozyme of the bacillus cereus S-12 strain product of CCTCC NO:M204089 by preserving number; Described filmogen is starch, dextrin, gelatin, agar, chitin or cellulose and derivant thereof; Described plasticizer is glycerol or sorbitol.
3,, it is characterized in that described component and weight portion are according to the pellicle of ocean lysozyme of claim 1
Ocean lysozyme 20.00
Filmogen 33.40
Plasticizer 54.7
Water 1670;
Described ocean lysozyme is to be the ocean lysozyme of the bacillus cereus S-12 strain product of CCTCC NO:M204089 by preserving number; Described filmogen is starch, dextrin, gelatin, agar, chitin or cellulose and derivant thereof; Described plasticizer is glycerol or sorbitol.
4, according to the pellicle of ocean lysozyme of claim 1 or 2 or 3, it is characterized in that
The physicochemical property of described ocean lysozyme is:
The molecular weight of lysozyme is 39KD;
The molecule of lysozyme is made up of 354 aminoacid;
The isoelectric point, IP of lysozyme is pI＞9;
The lysozyme optimum pH is 8;
The optimum temperature of lysozyme is 35 ℃;
Metal ion is to the influence of enzyme activity:
Most metal ion is lived to enzyme does not influence Co
2+, Cu
2+, Ag
+To enzyme slightly activation alive, Mg
2+, Zn
2+, Ca
2+Inhibitory action is arranged;
The pH stability and the heat stability of enzyme: enzyme is good stability between pH4.0-10.0, and enzyme is stable preferably in the time of 45 ℃;
Lysozyme reaches 3279.5u/mg than living behind the purification.
5,, it is characterized in that described filmogen is a hydroxypropyl emthylcellulose according to the pellicle of ocean lysozyme of claim 1 or 2 or 3; Plasticizer is a glycerol.
6, a kind of manufacture method of each pellicle of ocean lysozyme of claim 1-3 comprises the following steps:
(1) preparation
Filmogen is disperseed with boiling water, be stirred to room temperature, become transparent colloidal solution; With ocean lysozyme, with suitable quantity of water dissolving, the back adds in the filmogen colloidal solution, adds the plasticizer of above-mentioned formula ratio again, and adds water to Total Water and stir, and leaves standstill bubble collapse and gets colloidal solution;
(2) film
Step (1) colloidal solution is filmed, and wet film is dry under 38 ℃ of-40 ℃ of conditions, is divided into pellicle of ocean lysozyme behind the skinning.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510000033 CN1286525C (en) | 2005-01-05 | 2005-01-05 | New type pellicle of ocean lysozyme and preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200510000033 CN1286525C (en) | 2005-01-05 | 2005-01-05 | New type pellicle of ocean lysozyme and preparation method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1660418A CN1660418A (en) | 2005-08-31 |
| CN1286525C true CN1286525C (en) | 2006-11-29 |
Family
ID=35010155
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200510000033 Expired - Fee Related CN1286525C (en) | 2005-01-05 | 2005-01-05 | New type pellicle of ocean lysozyme and preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1286525C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101703093B (en) * | 2009-11-24 | 2013-02-13 | 青岛科谷生物制品研发有限公司 | Method for preparing novel ocean lysozyme antibiotic preservative coated film |
| CN102920685A (en) * | 2012-11-16 | 2013-02-13 | 沈阳药科大学 | Lysozyme film agent for eyes and preparation method thereof |
| CN103421329B (en) * | 2013-08-21 | 2015-07-01 | 华南理工大学 | Preparation method for antimicrobial edible film material |
-
2005
- 2005-01-05 CN CN 200510000033 patent/CN1286525C/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN1660418A (en) | 2005-08-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| TWI659696B (en) | Method for preparing feed additive containing high-concentration surface hormone | |
| CN108853473B (en) | Application of active octapeptide in protecting sensory hair cells | |
| CN1683399A (en) | Wasp antibacterial peptide and preparation method and application thereof | |
| CN111840516A (en) | Antibacterial gel for gynecology and preparation method thereof | |
| CN1309304C (en) | Sterilizing gel, preparing method and application thereof | |
| CN102405935B (en) | Protamine compounded preparation, preparation method and application thereof | |
| CN1286525C (en) | New type pellicle of ocean lysozyme and preparation method | |
| CN1704431A (en) | Melittin and use thereof | |
| CN102363040B (en) | Antimicrobial peptide preparation for mucosal tissues | |
| CN1484959A (en) | Sterilization formulation and wet tissue, preparation method and use | |
| US6992073B2 (en) | Use of N-acetyl-D-glucosamine in the manufacture of a medicament for treating cervical erosion | |
| CN100335126C (en) | Preparation for preventing and curing endometritis for dairy cattle and preparing method thereof | |
| CN1132517C (en) | Compound lysostaphin enzyme disinfectant | |
| CN1327741A (en) | Compound lysostaphin enzyme disinfectant | |
| CN1686165A (en) | Medical nano-carbon tube composition, preparation method and its application | |
| CN113024653A (en) | Pig-derived antibacterial peptide PMAP-23 variant and application thereof in preparation of feed | |
| CN1931170A (en) | Gatifloxacin ear drop and its prepn process | |
| CN1911441A (en) | Lysostaphin freeze dried powder used for preventing and treating cattle edometritis | |
| CN1164278C (en) | Biological preparation for treating burn and preparation method thereof | |
| CN1583169A (en) | Use of human lysozyme in preparation of medicine for hemorrhoid | |
| CN1911442A (en) | Lysostaphin freeze dried powder used for preventing and treating trauma surface infestation | |
| CN102178938A (en) | Marine lysozyme suppository and preparation method thereof | |
| CN1327047A (en) | Method for cultivating variotin pecilocin with silkworm and medicinal use | |
| CN1462584A (en) | Externally-used disinfector | |
| CN1279939C (en) | Medicinal spraying agent for treating nasitis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20061129 Termination date: 20150105 |
|
| EXPY | Termination of patent right or utility model |