CN109234256A - A kind of fermentation process of lysozyme - Google Patents
A kind of fermentation process of lysozyme Download PDFInfo
- Publication number
- CN109234256A CN109234256A CN201811346353.1A CN201811346353A CN109234256A CN 109234256 A CN109234256 A CN 109234256A CN 201811346353 A CN201811346353 A CN 201811346353A CN 109234256 A CN109234256 A CN 109234256A
- Authority
- CN
- China
- Prior art keywords
- column
- lysozyme
- added
- sodium
- fermentation process
- 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.)
- Pending
Links
Classifications
-
- 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)
- C12N9/2462—Lysozyme (3.2.1.17)
-
- 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)
- C12Y302/01017—Lysozyme (3.2.1.17)
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (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)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of fermentation process of lysozyme, step 1: dress column, cation exchange resin CM-Sephdex impregnates diel with dilute buffered saline solution, and be gently agitated for being allowed to balance frequently, with during this solvent balance, allow dress column CM-Sephdex material settle, the little particle floated in suspension is removed with decantation, then fill column, pay attention to not sandwiching bubble when filling column, column neglect process scale greatly depending on；Step 2: collecting extracting solution, collects egg white, after being filtered with double gauze to remove scrambled egg shell and embryonic cord, water is added by the doubling dose of its volume, careful stirring makes its mixing, it should be noted that is related to antalzyme fermentation technical field.The fermentation process of the lysozyme, is lysozyme finished product after being separated, dried by crystallization, and purity is insufficient, it is further purified by recrystallization method, after the attached solute of lysozyme is dissolved in solvent, after solvent is adsorbed, solute, which is carried out recrystallization, to be purified.
Description
Technical field
The present invention relates to antalzyme fermentation technical field, specially a kind of fermentation process of lysozyme.
Background technique
Lysozyme is also known as cytohydrolist, can act on bacteria cell wall peptide glycan molecule in specific manner, can make N- second
β-Isosorbide-5-Nitrae glycosidic bond fracture between acyl muramic acid and acetyl glucosamine ammonia, makes bacteria cell wall become loose and reaches dissolution carefully
The effect of bacterium.Lysozyme is distributed widely in high animal vegetable tissue and its secretion, protozoan, insect, plant and various
In microorganism.Lysozyme has multiple pharmacological effect as one of the nonspecific immune factors in human normal body fluid and tissue,
The reaction of body panimmunity is participated in, there is antibacterial, antiviral effect, in body normal defense function and nonspecific immunity, tool
There is the important function for keeping organism physiology balance.It can improve and enhance macrophage phagocytosis and digestive function, activate white thin
Born of the same parents' phagocytic function, and oligoleukocythemia caused by cytostatics can be improved, to enhance the resistance of body.Lysozyme energy
Direct hydrolysis gram-positive bacteria, secretory immunoglobulin A, complement participation under, moreover it is possible to hydrolyze Gram-negative bacteria such as
Escherichia coli etc..Equally there is bacteriolysis to drug tolerant bacteria, and there is spy significant in efficacy and to human body Small side effects
Point, thus be a kind of ideal medicinal enzyme.Lysozyme can directly be acted on negatively charged virus protein, with DNA, RNA,
Apoprotein forms double salt, makes virally inactivated.Lysozyme can participate in mucopolysaccharide metabolism, in cooperation as enzyme antimicrobial
While clothes and externally applied drug, strength antiinflammation can be played, it can also make its mistake in conjunction with the various acidic materials for inducing inflammation
It is living, and the curative effect of antibiotic and other medicines can be enhanced, improve the mucopolysaccharide metabolism of periplast, to reach anti-inflammatory, repair
The purpose of overlying tissue.Lysozyme is genetic engineering, cell engineering, essential toolenzyme in Fermentation Engineering.
Lysozyme is a kind of strong basicity albumen, and the crystal salt easy to form such as oxide, iodide and carbonate, in egg white
It is middle that a certain amount of above compound is added, and the pH value of solution is adjusted to its isoelectric point (PH=10.5), in optimum situation
Under, antalzyme crystallization will be precipitated at leisure, maximum output be reached in 72-96 hours, but existing egg white lysozyme is producing
When, it cannot be using egg yolk liquid when producing, so that yolk is largely wasted, and the egg white lysozyme purity of prior art production
It is not high, influence the quality of lysozyme.
Summary of the invention
(1) the technical issues of solving
In view of the deficiencies of the prior art, the present invention provides a kind of fermentation process of lysozyme, solve existing egg white
Lysozyme, cannot be using egg yolk liquid when producing, so that yolk is largely wasted, and the egg of prior art production in production
The problem of clear lysozyme purity is not high, influences lysozyme quality.
(2) technical solution
In order to achieve the above object, the present invention is achieved by the following technical programs: a kind of fermentation process of lysozyme, tool
Body the following steps are included:
Step 1: dress column, cation exchange resin CM-Sephdex impregnate diel with dilute buffered saline solution, and frequently
Be gently agitated for being allowed to balance, with during this solvent balance, allow dress column CM-Sephdex material settle, removed with decantation outstanding
The little particle floated in liquid, then fills column, pays attention to not sandwiching bubble when filling column, column neglect process scale greatly depending on；
Step 2: collecting extracting solution, collects egg white, after being filtered with double gauze to remove scrambled egg shell and embryonic cord, by it
Water is added in the doubling dose of volume, and careful stirring makes its mixing, it should be noted that speed is unsuitable too fast when stirring；Mixing direction
It must not change halfway；Cannot blister, splash bar should be smooth etc., otherwise will likely cause protein denaturation；
Step 3: PH precipitating is extremely slowly added into the hydrochloric acid of 0.5 mol/L into above-mentioned extracting solution, stirs when being added
It mixes, by the pH value adjustment of mixed liquor to 7.5 (noticing that hydrochloric acid should not be excessive), in a moment, in the filter tunnel for being placed with glass tampon
Middle this mixture of filtering gathers up filtrate, dense buffered saline melt is then added into filtrate to remove protein precipitation, measurement
The pH value of this solution is 8.2, such as if it is not, the hydrochloric acid or sodium hydroxide solution of view 0.5 mol/L of concrete condition are by pH value tune
To 8.2；
Step 4: aforesaid liquid is carefully added in CM-sephdex column by column chromatography, after extract enters in column,
First washed with suitable dilute buffer salt aqueous, then with it is aforementioned with PH=10.5 sodium carbonate -- sodium bicarbonate buffer liquid is washed
It is de-, until collecting eluent when lysozyme is all eluted from column；
Step 5: the above-mentioned eluent being collected into is adjusted its pH value with the sodium hydroxide solution of 0.5 mol/L by crystallization
It to after its isoelectric point, then is slowly added into common salt fine powder and makes total 0.3 mol/L of ultimate density, pay attention to stirring while adding,
Dissolve common salt in time, it is excessively high to avoid local salinity or be deposited on the bottom of container, after adding at low temperature (5 DEG C with
Under) stand it is allowed within 3--4 days to crystallize, such as crystallization formation completely after then sucking supernatant liquor, can be centrifugated out crystal.
Preferably, the molten wave of the dense buffer salt of sodium chloride-Tris-EDTA that the PH is 8.2 be added in every liter of water it is commercially available
Sodium chloride and each 0.5 mole of Tris-EDTA are formulated, dilute buffer salt aqueous by it is aforementioned with dense buffer salt aqueous add
10 times of water dilute and obtain.
Preferably, the hydrochloric acid solution and sodium hydroxide solution are the hydrochloric acid and sodium hydroxide solution of 0.5 mol/L, from
The hydrochloric acid of the sodium hydroxide and purity is high bought in chemical industry shop is formulated.
Preferably, the sodium bicarbonate buffer liquid is sodium carbonate -- the sodium bicarbonate buffer liquid of PH=10.5, from chemical industry quotient
The sodium carbonate and sodium bicarbonate that shop is bought are prepared, 0.8 mole of sodium carbonate and 0.2 mole of carbonic acid to be added in every liter of water
Hydrogen sodium forms.
Preferably, in the step 2, yolk can be used for extracting anti Bacillus pyocyaneu Flugge biochemical drug egg yolk oil or other use
On the way.
(3) beneficial effect
The present invention provides a kind of fermentation process of lysozyme.Have compared with prior art it is following the utility model has the advantages that
(1), the fermentation process of the lysozyme by dress column, collects extracting solution, PH precipitating, column chromatography and crystallization, cation
Exchanger resin CM-Sephdex impregnates diel with dilute buffered saline solution, and is gently agitated for being allowed to balance frequently, molten with this
During agent balances, allows the CM-Sephdex material of dress column to settle, the little particle floated in suspension is removed with decantation, is then filled
Column pays attention to not sandwiching bubble when filling column, column neglect process scale greatly depending on；Collect egg white, with double gauze filter with
After removing scrambled egg shell and embryonic cord, water is added by the doubling dose of its volume, careful stirring makes its mixing, it should be noted that stirring
Shi Sudu is unsuitable too fast；Mixing direction must not change halfway；Cannot blister, splash bar should be smooth etc., otherwise will likely cause egg
White matter denaturation；It is extremely slowly added into the hydrochloric acid of 0.5 mol/L into above-mentioned extracting solution, stirs while adding, by mixed liquor
PH value adjustment in a moment, filters this mixture to 7.5 (noticing that hydrochloric acid should not be excessive) in the filter tunnel for being placed with glass tampon
To remove protein precipitation, filtrate is gathered up, dense buffered saline melt is then added into filtrate, the pH value for measuring this solution is
8.2, such as if it is not, pH value is adjusted to 8.2 with the hydrochloric acid of 0.5 mol/L or sodium hydroxide solution depending on concrete condition；Carefully will
Aforesaid liquid is added in CM-sephdex column, after extract enters in column, is first washed with suitable dilute buffer salt aqueous, is then used
It is aforementioned with PH=10.5 sodium carbonate -- sodium bicarbonate buffer liquid elution, collect eluent until lysozyme is all from column
Until when elution；The above-mentioned eluent being collected into is adjusted into its pH value to its isoelectric point with the sodium hydroxide solution of 0.5 mol/L
Afterwards, then be slowly added into common salt fine powder and make total 0.3 mol/L of ultimate density, pay attention to stirring while adding, make common salt and
When dissolve, excessively high to avoid local salinity or be deposited on the bottom of container, (5 DEG C or less) standing 3--4 at low temperature after adding
It allows it to crystallize, and such as crystallization is formed completely, then after sucking supernatant liquor, can be centrifugated out crystal；By crystallization through dividing
From, it is dry after be lysozyme finished product, purity is insufficient, is further purified by recrystallization method, by the attached solute of lysozyme
After being dissolved in solvent, after solvent is adsorbed, solute, which is carried out recrystallization, to be purified.
(2), the fermentation process of the lysozyme is collected egg white, is filtered with double gauze to remove by collecting extracting solution
After scrambled egg shell and embryonic cord, water is added by the doubling dose of its volume, careful stirring makes its mixing, it should be noted that stirring speed per hour
Degree is unsuitable too fast；Mixing direction must not change halfway；Cannot blister, splash bar should be smooth etc., otherwise will likely cause protein
Denaturation, yolk can be used for extracting anti Bacillus pyocyaneu Flugge biochemical drug egg yolk oil or other purposes, solve existing egg white lysozyme
Enzyme is in production, egg yolk liquid when cannot be using production so that the appearance for the case where yolk is largely wasted, can by yolk into
Row utilizes, to produce other products, increased economic benefit, the income of Ti Ge factory.
Specific embodiment
Technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is only
It is a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill people
Member's every other embodiment obtained without making creative work, shall fall within the protection scope of the present invention.
The embodiment of the present invention provides a kind of technical solution: a kind of fermentation process of lysozyme, specifically includes the following steps:
Step 1: dress column, cation exchange resin CM-Sephdex impregnate diel with dilute buffered saline solution, and frequently
Be gently agitated for being allowed to balance, with during this solvent balance, allow dress column CM-Sephdex material settle, removed with decantation outstanding
The little particle floated in liquid, then fills column, pays attention to not sandwiching bubble when filling column, column neglect process scale greatly depending on；
Step 2: collecting extracting solution, collects egg white, after being filtered with double gauze to remove scrambled egg shell and embryonic cord, by it
Water is added in the doubling dose of volume, and careful stirring makes its mixing, it should be noted that speed is unsuitable too fast when stirring；Mixing direction
It must not change halfway；Cannot blister, splash bar should be smooth etc., otherwise will likely cause protein denaturation；
Step 3: PH precipitating is extremely slowly added into the hydrochloric acid of 0.5 mol/L into above-mentioned extracting solution, stirs when being added
It mixes, by the pH value adjustment of mixed liquor to 7.5 (noticing that hydrochloric acid should not be excessive), in a moment, in the filter tunnel for being placed with glass tampon
Middle this mixture of filtering gathers up filtrate, dense buffered saline melt is then added into filtrate to remove protein precipitation, measurement
The pH value of this solution is 8.2, such as if it is not, the hydrochloric acid or sodium hydroxide solution of view 0.5 mol/L of concrete condition are by pH value tune
To 8.2；
Step 4: aforesaid liquid is carefully added in CM-sephdex column by column chromatography, after extract enters in column,
First washed with suitable dilute buffer salt aqueous, then with it is aforementioned with PH=10.5 sodium carbonate -- sodium bicarbonate buffer liquid is washed
It is de-, until collecting eluent when lysozyme is all eluted from column；
Step 5: the above-mentioned eluent being collected into is adjusted its pH value with the sodium hydroxide solution of 0.5 mol/L by crystallization
It to after its isoelectric point, then is slowly added into common salt fine powder and makes total 0.3 mol/L of ultimate density, pay attention to stirring while adding,
Dissolve common salt in time, it is excessively high to avoid local salinity or be deposited on the bottom of container, after adding at low temperature (5 DEG C with
Under) stand it is allowed within 3--4 days to crystallize, such as crystallization formation completely after then sucking supernatant liquor, can be centrifugated out crystal.
The molten wave of the dense buffer salt of sodium chloride-Tris-EDTA that PH is 8.2 be added in every liter of water commercially available sodium chloride and
Each 0.5 mole of Tris-EDTA is formulated, dilute buffer salt aqueous by it is aforementioned with dense buffer salt aqueous add 10 times of water dilution and
, hydrochloric acid solution and sodium hydroxide solution are the hydrochloric acid and sodium hydroxide solution of 0.5 mol/L, are bought from chemical industry shop
Sodium hydroxide and the hydrochloric acid of purity is high are formulated, and sodium bicarbonate buffer liquid is sodium carbonate -- the sodium bicarbonate buffer of PH=10.5
Liquid, the sodium carbonate and sodium bicarbonate bought from chemical industry shop are prepared, in every liter of water be added 0.8 mole sodium carbonate and
0.2 mole of sodium bicarbonate forms, and in step 2, yolk can be used for extracting the biochemical drug egg yolk oil or other of anti Bacillus pyocyaneu Flugge
Purposes.
It should be noted that, in this document, relational terms such as first and second and the like are used merely to a reality
Body or operation are distinguished with another entity or operation, are deposited without necessarily requiring or implying between these entities or operation
In any actual relationship or order or sequence.Moreover, the terms "include", "comprise" or its any other variant are intended to
Non-exclusive inclusion, so that the process, method, article or equipment including a series of elements is not only wanted including those
Element, but also including other elements that are not explicitly listed, or further include for this process, method, article or equipment
Intrinsic element.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with
A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding
And modification, the scope of the present invention is defined by the appended.
Claims (5)
1. a kind of fermentation process of lysozyme, it is characterised in that: specifically includes the following steps:
Step 1: dress column, cation exchange resin CM-Sephdex impregnate diel with dilute buffered saline solution, and frequently gently
Agitation be allowed to balance, with during this solvent balance, allow dress column CM-Sephdex material settle, removed in suspension with decantation
The little particle of floating, then fills column, pays attention to not sandwiching bubble when filling column, column neglect process scale greatly depending on；
Step 2: collecting extracting solution, collects egg white, after being filtered with double gauze to remove scrambled egg shell and embryonic cord, by its volume
Doubling dose water is added, careful stirring makes its mixing, it should be noted that speed is unsuitable too fast when stirring；Mixing direction must not
Midway changes；Cannot blister, splash bar should be smooth etc., otherwise will likely cause protein denaturation；
Step 3: PH precipitating is extremely slowly added into the hydrochloric acid of 0.5 mol/L into above-mentioned extracting solution, stirs while adding,
By the pH value adjustment of mixed liquor to 7.5 (noticing that hydrochloric acid should not be excessive), in a moment, the mistake in the filter tunnel for being placed with glass tampon
This mixture is filtered to remove protein precipitation, gathers up filtrate, dense buffered saline melt is then added into filtrate, it is molten to measure this
The pH value of liquid is 8.2, such as if it is not, being adjusted to pH value with the hydrochloric acid of 0.5 mol/L or sodium hydroxide solution depending on concrete condition
8.2；
Step 4: aforesaid liquid is carefully added in CM-sephdex column, after extract enters in column, first uses by column chromatography
Suitable dilute buffer salt aqueous is washed, and then with the sodium carbonate of the aforementioned PH=10.5 matched -- sodium bicarbonate buffer liquid elutes, is received
Until collecting eluent when lysozyme is all eluted from column；
Step 5: the above-mentioned eluent being collected into is adjusted its pH value with the sodium hydroxide solution of 0.5 mol/L and arrives it by crystallization
It after isoelectric point, then is slowly added into common salt fine powder and makes total 0.3 mol/L of ultimate density, pay attention to stirring while adding, make chlorine
Timely dissolution is received in change, excessively high to avoid local salinity or be deposited on the bottom of container, and (5 DEG C or less) are quiet at low temperature after adding
It sets and it is allowed within 3--4 days to crystallize, such as crystallization is formed completely, then after sucking supernatant liquor, can be centrifugated out crystal.
2. a kind of fermentation process of lysozyme according to claim 1, it is characterised in that: the chlorination that the PH is 8.2
The molten wave of the dense buffer salt of sodium-Tris-EDTA be added in every liter of water each 0.5 mole of commercially available sodium chloride and Tris-EDTA prepare and
At dilute buffer salt aqueous is added 10 times of water dilutions by the aforementioned dense buffer salt aqueous matched and obtained.
3. a kind of fermentation process of lysozyme according to claim 1, it is characterised in that: the hydrochloric acid solution and hydroxide
Sodium solution is the hydrochloric acid and sodium hydroxide solution of 0.5 mol/L, the salt of the sodium hydroxide bought from chemical industry shop and purity is high
Acid is formulated.
4. a kind of fermentation process of lysozyme according to claim 1, it is characterised in that: the sodium bicarbonate buffer liquid is
The sodium carbonate of PH=10.5 -- sodium bicarbonate buffer liquid, the sodium carbonate and sodium bicarbonate bought from chemical industry shop are prepared, with
0.8 mole of sodium carbonate is added in every liter of water and 0.2 mole of sodium bicarbonate forms.
5. a kind of fermentation process of lysozyme according to claim 1, it is characterised in that: in the step 2, yolk can
For extract anti Bacillus pyocyaneu Flugge biochemical drug egg yolk oil or other purposes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811346353.1A CN109234256A (en) | 2018-11-13 | 2018-11-13 | A kind of fermentation process of lysozyme |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811346353.1A CN109234256A (en) | 2018-11-13 | 2018-11-13 | A kind of fermentation process of lysozyme |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109234256A true CN109234256A (en) | 2019-01-18 |
Family
ID=65078295
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811346353.1A Pending CN109234256A (en) | 2018-11-13 | 2018-11-13 | A kind of fermentation process of lysozyme |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109234256A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117050854A (en) * | 2023-07-14 | 2023-11-14 | 江苏天成蛋业有限公司 | Preparation device for extracting lysozyme from egg white and application method thereof |
-
2018
- 2018-11-13 CN CN201811346353.1A patent/CN109234256A/en active Pending
Non-Patent Citations (1)
| Title |
|---|
| 施德恒: "一种溶菌酶的提取方法", 《今日科技》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117050854A (en) * | 2023-07-14 | 2023-11-14 | 江苏天成蛋业有限公司 | Preparation device for extracting lysozyme from egg white and application method thereof |
| CN117050854B (en) * | 2023-07-14 | 2024-07-23 | 江苏天成蛋业有限公司 | Preparation device for extracting lysozyme from egg white and application method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103045707B (en) | Method for preparing rice protein active polypeptide through continuous enzymolysis and secondary membrance filtering and purification | |
| CN110028533A (en) | A kind of method and application of the refining amino glucosamine salt hydrochlorate from microbial fermentation solution | |
| CN109234256A (en) | A kind of fermentation process of lysozyme | |
| CH621769A5 (en) | ||
| CN109053508A (en) | A kind of preparation method of carbocisteine | |
| JP5908496B2 (en) | Method for extracting human serum albumin from the grain of transgenic rice | |
| FI102620B (en) | Process for decolorizing solutions | |
| CN107760740A (en) | A kind of big sugar of high-purity kanjak mannan and preparation method thereof, application | |
| EP2208734B1 (en) | Food material for inhibiting the formation of osteoclast | |
| CN100467061C (en) | Scorpionfish-ink polysaccharide and its preparation | |
| CN101700897B (en) | Pharmaceutical grade anhydrous sodium carbonate and preparation method thereof | |
| JPH0569116B2 (en) | ||
| CN104419742B (en) | A kind of method that gelatin is prepared by raw material of skeletal grain | |
| CN104371910A (en) | Method for extracting collagen from deer bones | |
| CN107746868A (en) | The preparation method of isomaltooligosaccharide syrup | |
| CN104628766B (en) | Industrial manufacturing method of glycerol phosphocholine | |
| AU2008320272B2 (en) | Food material for inhibiting bone resorption | |
| CN103554136A (en) | Preparation method of cefmenoxine hydrochloride dry powder | |
| CN107836613A (en) | Chlorella growth factor spirulina polysaccharide drink with function preparation method | |
| CN104593345A (en) | Method for separation and purification of trypsin and pharmaceutical composition containing trypsin | |
| CN106729615A (en) | A kind of antibacterial biological film and preparation method thereof | |
| CN207468525U (en) | A kind of active compound purification system of Cefixime granule production technology section | |
| CN105767997A (en) | Bee pollen and preparation method thereof | |
| CN102382150B (en) | Preparation method of high-purity mixed sodium deoxyribonucleotide | |
| CN204211727U (en) | A kind of system extracting collagen protein from deer bone |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190118 |
|
| RJ01 | Rejection of invention patent application after publication |