CN106632646A - Method of co-purifing three microfunctional proteins from eggshell - Google Patents
Method of co-purifing three microfunctional proteins from eggshell Download PDFInfo
- Publication number
- CN106632646A CN106632646A CN201710030574.7A CN201710030574A CN106632646A CN 106632646 A CN106632646 A CN 106632646A CN 201710030574 A CN201710030574 A CN 201710030574A CN 106632646 A CN106632646 A CN 106632646A
- Authority
- CN
- China
- Prior art keywords
- eggshell
- egg
- kinds
- copurification
- exchange chromatography
- 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
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/465—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from birds
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a method of co-purifing three microfunctional proteins from eggshell. A buffer liquid is adopted as an eggshell protein extractant, and a Yin-Yang ion exchange chromatography column technology technology is adopted to conduct enrichment, separation and purification to obtain three microfunctional proteins relevant to biomineralization only one time. The invention solves the problems of difficult eggshell protein extraction and enrichment, low efficiency and the like, and builds a simple, quick and efficient copurification method; besides the three proteins have high-purity (>90%), the overall process takes short time, the production cost is low, and the operation is simple; the method of co-purifing the three microfunctional proteins from the eggshell has wide industrialized production potential, and provides the research and application of functional protein in the eggshell with advantages.
Description
Technical field
The present invention relates to protein purification preparing technical field, in particular to one kind from eggshell three kinds of micro work(of copurification
The method of energy albumen.
Background technology
At present, more, such as chorion is only found in eggshell with the protein of critical function such as biomineralization and crystallizations
Mineralization protein -17 (Ovocleidin-17, OC-17), chorion reinforcing protein-11 6 (Ovocleidin-116, OC-116) and ovum
Shell fat hop protein -36 (Ovocalyxin-36, OCX-36) etc., these protein not only have remarkable biomineralization with crystallization
Function, also with bioconjugation, catalysis, enzyme modulation, biomineralization, antibacterial, molecule conduction and structural molecule transhipment, passage regulation and control
Deng critical function, there is extremely wide application prospect in the field such as biology, medicine, food, material.
But these albumen content in eggshell seldom, is studied and shows that OC-17, OC-116 and OCX-36 are primarily present in eggshell
In, the 0.4 ‰ of Eggshell weight, 0.8 ‰, 1 ‰ are accounted for respectively, because content is extremely low, extract very difficult, enrichment method difficulty is very
Height, and traditional acidity extraction eggshell albumen can cause different degrees of damage to protein active, thus, it is necessary to develop one kind
Leniently extracting method.In addition, current eggshell protein purification is separated only by the single albumen of purifying, rarely combine both at home and abroad
The research report of multiple protein is extracted, therefore there is many defects such as waste eggshell resource, extraction efficiency is low, energy loss is big,
So as to further limit these albumen Quality Research applications.Once breaking through above technical bottleneck, can not only make full use of extremely
Abundant eggshell resource, in fields such as biological medicine, material, light industry, functional food huge commercial application value is played, moreover it is possible to
The discarded pollution to environment of eggshell is prevented, with great economic and social benefits.
The content of the invention
The purpose of the present invention is for defect present in above prior art, there is provided one kind combined extracting three from eggshell
The method for planting micro functional protein.
To achieve these goals, the invention provides a kind of side of three kinds of micro functional proteins of the combined extracting from eggshell
Method, comprises the following steps：
1) fresh eggshell is cleaned up, and is crushed after drying, obtains egg-shell meal,
2) egg-shell meal and water are mixed and stirred for, collect lower sediment, the egg-shell meal for sloughing egg shell membrane is obtained final product after drying；
3) to step 2) gained sloughs and add in the egg-shell meal of egg shell membrane neutral buffered liquid and extract, stir 10～30 hours,
Supernatant is collected, eggshell protein crude extract is obtained final product；
4) anion exchange chromatography, cation-exchange chromatography post are connected, by step 3) obtained by supernatant on
Post, with equilibrium liquid 1～3 times of column volume is eluted, and eggshell protein is combined with the filler in ion exchange column；
5) anion exchange chromatography and cation-exchange chromatography post of series connection are taken apart, is eluted respectively and collected and washed
De- peak；
6) each eluting peak collected is carried out into dialysis desalting and freeze-drying, wherein, cation-exchange chromatography post is washed
De- peak is Ovocleidin-17, and first eluting peak of anion exchange chromatography is Ovocalyxin-36, second
Eluting peak is Ovocleidin-116.
Further, the step 1) in, the temperature of baking is 25～35 DEG C, and the time is 12～24h, eggshell grinding particle size
For 50～300 mesh.
Yet further, the step 2) in, egg-shell meal is 1 with the mass ratio of water:5～20.
Yet further, the step 3) in, the neutral buffered extract is phosphate buffer.
Yet further, the step 3) in, egg-shell meal is 1 with the mass ratio of phosphate buffer:5～20, phosphate
PH of cushioning fluid 5.5～8.5.
Yet further, the step 3) in, stirring extraction time is 24～32h.
Yet further, the step 4) in, equilibrium liquid is the Tris-HCl of 50～200mM, pH 6.8～8.8.
Yet further, the step 5) in, the eluent of anion exchange chromatography is pH value 6.2～7.8, salinity
The buffer solution of 0.1～0.5mol/L, 0.5～5mmol/L of reducing agent.
Yet further, the step 5) in, the eluent of cation-exchange chromatography is pH value 6.8～8.8, salinity
The buffer solution of 0.1～0.5mol/L, 0.5～5mmol/L of reducing agent.
Yet further, the reducing agent is dithiothreitol (DTT) or beta -mercaptoethanol.
The inventive method condition is chosen
1) selection of Pretreatment
(1) eggshell grinding particle size test：50 mesh, 100 mesh, 200 mesh, the process of 300 mesh granularities are set, and other conditions are mutually sympathized with
Under condition, relatively more each eggshell rate of recovery for processing, variation tendency is shown in figure.Granularity has the higher eggshell rate of recovery in 100 mesh,
In consideration of it, egg-shell meal selects 100 mesh granularities (as shown in Figure 2).
(2) solid-liquid ratio test：It is 1 that egg-shell meal is arranged with water solid-liquid ratio:5、1:10、1:15、1:20, other operating conditions
Under, relatively more each eggshell rate of recovery for processing, variation tendency such as Fig. 3.Solid-liquid ratio 1:Eggshell has the higher rate of recovery when 10.
2) selection of protein extracting method condition
(1) selection of solution is extracted：It is different except solution is extracted, extract protein, Detection and Extraction liquid in same procedure condition
In protein content, and according to below equation calculate extraction rate of protein.
Protein (g)/egg-shell meal quality (g) × 100% in extraction rate of protein (%)=extract
Final recovery rate is as shown in the table, compares for convenience, and result is also listed in the lump.
Impact of the solution of table 1 to recovery rate
| Extract solution | PH value | Extraction rate of protein |
| Phosphate buffer | 6.8 | 2.35% |
| Acetate buffer | 4.7 | 1.67% |
| Citrate buffer solution | 5.5 | 1.85% |
| Tris buffer solutions | 7.6 | 1.43% |
After testing, the content of protein is about 2.63% in egg-shell meal, and the result of table 1 shows and adopts phosphate-buffered simultaneously
Liquid energy enough extracts 89.7% eggshell protein.
(2) solid-liquid ratio test：Egg-shell meal is set：Extract is 1:5、1:10、1:15、1:20 are tested, except solid-liquid ratio
Example is different, to extract protein, the protein content in Detection and Extraction liquid with the same procedure condition of embodiment 1, and calculates egg
White matter recovery rate, as a result as shown in Figure 4.Solid-liquid ratio is 1:10 can extract higher eggshell albumen, therefore select solid-liquid ratio
For 1:10.
(3) extraction time test：Extraction time 8h, 16h, 24h, 32h, the Protein Extraction of each process time of comparison are set
Rate, as a result as shown in figure 5, the yield of 32h is higher, then extends the time, increases change not substantially, set extraction time as 24～
32h。
The beneficial effects of the present invention is：
(1) present invention establishes a kind of isolating and purifying for biomineralization, crystallization and its quality crystallization control related protein
Method, the method mild condition is conducive to keeping the native conformation and activity of functional protein；
(2) present invention only needs to a separating step and the control of various biomineralizations, crystallization and its quality crystallization is obtained
Related functional protein；
(3) present invention is enriched with, ion according to the characteristic of three kinds of micro functional proteins using anions and canons displacement chromatography post
Displacement chromatography technology is easily enlarged metaplasia product, therefore, the technology has the potentiality for being applied to industrialized production；
(4) the extremely micro biomineralization of three kinds of eggshells of the invention, crystallization and its quality crystallization control function albumen, have
Fractional dose is big, yield is high, purity is high, environmental protection, integrated cost are low, it is efficiently quick the characteristics of.
Description of the drawings
Fig. 1 for the present invention from egg shell combined extracting Ovocleidin-17, Ovocleidin-116 and
The method flow schematic diagram of Ovocalyxin-36；
Fig. 2 is impact of the eggshell Powder Particle Size to the rate of recovery；
Fig. 3 is impact of the solid-liquid ratio of egg-shell meal and water to the rate of recovery；
Fig. 4 is impact of the solid-liquid ratio of egg-shell meal and extract to extraction rate of protein；
Fig. 5 is mixing time to the impact to extraction rate of protein.
Fig. 6 is anion-exchange chromatography collection of illustrative plates；
Fig. 7 is cation-exchange chromatography collection of illustrative plates；
Fig. 8 is the SDS- acrylamide gel electrophoresis figures at each peak of Jing anion-exchange chromatographies gained；
Fig. 9 is the SDS- acrylamide gel electrophoresis figures at Jing cation-exchange chromatographies gained peak；
Figure 10 is the electrospray ionization mass spectrum figure of Ovocleidin-116；
Figure 11 is the HPLC collection of illustrative plates of Ovocalyxin-36；
Figure 12 is the HPLC collection of illustrative plates of Ovocleidin-17；
Figure 13 is the HPLC collection of illustrative plates of Ovocleidin-116.
In wherein Fig. 8,1,2 are respectively Ovocalyxin-36 and Ovocleidin-116.
Specific embodiment
In order to preferably explain the present invention, the main contents of the present invention are further elucidated below in conjunction with specific embodiment, but
Present disclosure is not limited solely to following examples.
Embodiment 1
(1) sample pre-treatments
Fresh egg shell is cleaned up, drying, crushes 200 mesh sieves, and with water 1 is pressed：10 mixing, stir 15min, lower floor
Suction filtration drying obtains the egg-shell meal handled well；
(2) Protein Extraction
The egg-shell meal that 100g is handled well is sufficiently stirred in pH 6.8,1000mL 2mM phosphate buffers, magnetic agitation
24h, then at 4 DEG C, is centrifuged 15min under conditions of 12000 × g, take supernatant, obtains eggshell protein crude extract.
(3) ion exchange column of balance series connection
The Tris-HCl of 50mM, pH 8.5, with the CM ion exchange columns of the flow velocity balance series connection of 1mL/min, (GE is public
Department's production, 17018001) with Q Sepharose ion exchange columns (GE companies produce, article No. 17051010).
(4) protein solution upper prop
The protein crude extract injection chromatographic column that step (2) is obtained, eluent is Tris-HCl or PBS, is contained
The reducing agent of 0.5mM, pH is 8.8, elutes one times of column volume.
(5) anion exchange chromatography is eluted
Remove CM ion exchange columns, eluent is Na-Hepe buffer solutions, pH 8.2, the sodium chloride containing 100mM,
The reducing agent of 5mM.Eluted with the flow velocity of 1mL/min, collected eluting peak.
(6) cation-exchange chromatography post is eluted
With pH 8.5, the solution containing 50mM Tris-HCl and 400mM sodium chloride is eluent, with the flow velocity of 1mL/min
Eluted, collected eluting peak.
Embodiment 2
(1) sample pre-treatments
Fresh egg shell is cleaned up, drying, crushes 200 mesh sieves, and with water 1 is pressed：10 mixing, stir 15min, lower floor
Suction filtration drying obtains the egg-shell meal handled well；
(2) Protein Extraction
The egg-shell meal that 100g is handled well is sufficiently stirred in pH 6.8,1000mL 10mM phosphate buffers, magnetic agitation
28h, then at 4 DEG C, is centrifuged 15min under conditions of 12000 × g, take supernatant, obtains eggshell protein crude extract.
(3) ion exchange column of balance series connection
The Tris-HCl of 50mM, pH 8.5, with the CM ion exchange columns of the flow velocity balance series connection of 1mL/min, (GE is public
Department's production, 17018001) with DEAE ion exchange columns (GE companies produce, article No. 17070901).(4) protein solution upper prop
The protein crude extract injection chromatographic column that step (2) is obtained, eluent is Tris-HCl or PBS, the reduction containing 2mM
Agent, pH is 8.0, elutes one times of column volume.
(5) anion exchange chromatography is eluted
Remove CM ion exchange columns, eluent is Na-Hepe buffer solutions, pH 7.0, the sodium chloride containing 200mM,
The reducing agent of 2mM.Eluted with the flow velocity of 1mL/min, collected eluting peak.
(6) cation-exchange chromatography post pH 8.5, the solution containing 100mM Tris-HCl and 300mM sodium chloride are eluted
For eluent, eluted with the flow velocity of 1mL/min, collected eluting peak.
Embodiment 3
(1) sample pre-treatments
Fresh egg shell is cleaned up, drying, crushes 100 mesh sieves, and with water 1 is pressed：10 mixing, stir 15min, lower floor
Suction filtration drying obtains the egg-shell meal handled well；
(2) Protein Extraction
The egg-shell meal that 100g is handled well is sufficiently stirred in pH 6.8,1000mL 5mM phosphate buffers, magnetic agitation
32h, then at 4 DEG C, is centrifuged 15min under conditions of 12000 × g, take supernatant, obtains eggshell protein crude extract.
(3) ion exchange column of balance series connection
The Tris-HCl of 50mM, pH 8.5, with the SP Sepharose ion exchange layers of the flow velocity balance series connection of 1mL/min
(GE companies produce analysis post, 17072910) with DEAE ion exchange columns (GE companies produce, article No. 17070901).
(4) protein solution upper prop
The protein crude extract injection chromatographic column that step (2) is obtained, eluent is Tris-HCl or PBS, containing 5mM
Reducing agent, pH is 7.2, elute one times of column volume.
(5) anion exchange chromatography is eluted
Remove CM ion exchange columns, eluent is Na-Hepe buffer solutions, pH 6.8, the sodium chloride containing 500mM,
The reducing agent of 0.5mM.Eluted with the flow velocity of 1mL/min, collected eluting peak.
(6) cation-exchange chromatography post is eluted
With pH 8.5, the solution containing 50mM Tris-HCl and 200mM sodium chloride is eluent, with the flow velocity of 1mL/min
Eluted, collected eluting peak.
Embodiment 4
(1) SDS- polyacrylamide gel electrophoresises identification
The μ L of eluting peak sample solution 80 collected after chromatography are taken, 20 μ L albumen sample-loading buffers are added, is mixed, boiling water bath adds
Hot 5min, 10 μ L are taken after cooling carries out conventional SDS-PAGE electrophoresis, resolving gel concentration 12%, constant voltage 100V.Electrophoresis result
As shown in Figure 8,9.Swimming lane " 0 " sample in Fig. 8 is eggshell crude extract, shows several electrophoretic bands, shows that eggshell albumen is thick
Extract is the mixture of multiple proteins.Fig. 8 swimming lanes " 1 " and " 2 " correspond to respectively first peak and second peak in Fig. 6, point
Son amount is respectively 100-110kDa and 34-37kDa, bibliography report and Protein Data Bank data, with its molecule in eggshell
The consistent protein of amount is respectively Ovocalyxin-36 and Ovocleidin-116.The chromatography collection of illustrative plates of Fig. 9 correspondence Fig. 7, molecular weight
It is Ovocleidin-17 for 15-18kDa.
(2) purity testing
Using high effective liquid chromatography for measuring purity, to integrating peak areas, according to peak area ratio to each protein product
Purity is quantitatively calculated, and as shown in Figure 11～13, as a result be see the table below：
As described above, using the invention provides one kind from egg shell with technique copurification Ovocleidin-17,
The method of Ovocleidin-116 and Ovocalyxin-36, it is possible to obtain the higher product of higher protein extracting ratio and purity.
Other unspecified parts are prior art.Although above-described embodiment is made that to the present invention retouching in detail
State, but it is only a part of embodiment of the invention, rather than whole embodiments, people can with according to the present embodiment without
Other embodiment is obtained under the premise of creativeness, these embodiments belong to the scope of the present invention.
Claims (10)
1. a kind of method of three kinds of micro functional proteins of the copurification from eggshell, it is characterised in that：Comprise the following steps：
1) fresh eggshell is cleaned up, and is crushed after drying, obtains egg-shell meal,
2) egg-shell meal and water are mixed and stirred for, collect lower sediment, the egg-shell meal for sloughing egg shell membrane is obtained final product after drying；
3) to step 2) gained sloughs and add in the egg-shell meal of egg shell membrane neutral buffered liquid and extract, stir 10～30 hours, collect
Supernatant is simultaneously dialysed, and obtains final product eggshell protein crude extract；
4) anion exchange chromatography, cation-exchange chromatography post are connected, by step 3) obtained by supernatant upper prop, use
Equilibrium liquid elutes 1～3 times of column volume, eggshell protein is combined with the filler in ion exchange column；
5) anion exchange chromatography and cation-exchange chromatography post of series connection are taken apart, is eluted and collected wash-out respectively
Peak；
6) each eluting peak collected is carried out into dialysis desalting and freeze-drying, wherein, the eluting peak of cation-exchange chromatography post
As Ovocleidin-17, first eluting peak of anion exchange chromatography is Ovocalyxin-36, second wash-out
Peak is Ovocleidin-116.
2. according to claim 1 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 1) in, the temperature of baking is 25～35 DEG C, and the time is 12～24h, and eggshell grinding particle size is 50～300 mesh.
3. the method for three kinds of micro functional proteins of the copurification from eggshell according to claim 1 or claim 2, it is characterised in that：Institute
State step 2) in, egg-shell meal is 1 with the mass ratio of water:5～20.
4. the method for three kinds of micro functional proteins of the copurification from eggshell according to claim 1 or claim 2, it is characterised in that：Institute
State step 3) in, the neutral buffered extract is phosphate buffer.
5. according to claim 4 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 3) in, the mass ratio of egg-shell meal and phosphate buffer is 1:5～20, phosphate buffer pH value 5.5～8.5.
6. according to claim 5 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 3) in, stirring extraction time be 24～32h.
7. according to claim 1 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 4) in, equilibrium liquid is the Tris-HCl of 50～200mM, pH 6.8～8.8.
8. according to claim 1 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 5) in, the eluent of anion exchange chromatography be pH value 6.2～7.8,0.1～0.5mol/L of salinity, reducing agent 0.5～
The buffer solution of 5mmol/L.
9. according to claim 8 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：The step
It is rapid 5) in, the eluent of cation-exchange chromatography be pH value 6.8～8.8,0.1～0.5mol/L of salinity, reducing agent 0.5～
The buffer solution of 5mmol/L.
10. according to claim 8 or claim 9 from eggshell three kinds of micro functional proteins of copurification method, it is characterised in that：Institute
State step 5) in, reducing agent is dithiothreitol (DTT) or beta -mercaptoethanol.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710030574.7A CN106632646A (en) | 2017-01-16 | 2017-01-16 | Method of co-purifing three microfunctional proteins from eggshell |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710030574.7A CN106632646A (en) | 2017-01-16 | 2017-01-16 | Method of co-purifing three microfunctional proteins from eggshell |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106632646A true CN106632646A (en) | 2017-05-10 |
Family
ID=58841931
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710030574.7A Pending CN106632646A (en) | 2017-01-16 | 2017-01-16 | Method of co-purifing three microfunctional proteins from eggshell |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106632646A (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875669A (en) * | 2012-10-29 | 2013-01-16 | 天津商业大学 | Method for separating and extracting ovotransferrin |
-
2017
- 2017-01-16 CN CN201710030574.7A patent/CN106632646A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102875669A (en) * | 2012-10-29 | 2013-01-16 | 天津商业大学 | Method for separating and extracting ovotransferrin |
Non-Patent Citations (2)
| Title |
|---|
| MAOJIE ZHANG ET AL.: "An Efficient Method for Co-purification of Eggshell Matrix Proteins OC-17, OC-116, and OCX-36", 《KOREAN J. FOOD SCI. AN.》 * |
| 张茂杰等: "禽蛋壳基质蛋白质ovocalyxin-36在蛋壳中的分布、基因结构以及生物活性", 《动物营养学报》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102952187B (en) | Preparation method of high-purity bovine serum albumin | |
| US8574883B2 (en) | Method of co-extracting multiple proteins from chicken egg white | |
| Tiselius et al. | Separation and Fractionation of Macromolecules and Particles: New or improved separation procedures underlie many important recent advances in biochemistry. | |
| CN104292327A (en) | Method for extracting phycobiliprotein from spirulina | |
| CN105624251B (en) | A kind of peony seeds blood sugar reducing peptide and its purification process and application | |
| CN107216228B (en) | Eutectic solvent and method for extracting anthraquinone in rheum officinale | |
| CN104759269B (en) | Preparation method of graphene microsphere biological separation medium with controllable particle size | |
| CN107840883B (en) | Method for simultaneously separating 3 main components of bovine milk casein | |
| CN105017412A (en) | Method for separating high-purity bovine serum albumin from bovine serum | |
| CN108558971A (en) | A kind of preparation method of roselle anthocyanin | |
| CN104926680A (en) | Method for extracting bio-active substances from low-grade tea and tea-manufacturing leftovers | |
| CN105821093A (en) | Lactobacillus plantarum exopolysaccharide and preparation method thereof | |
| CN102702341B (en) | Recombinant human nerve growth factor purifying method based on CHO cell expression system | |
| Janson | Columns for large-scale gel filtration on porous gels. Fractionation of rape seed proteins and insulin | |
| CN109879930B (en) | Purification method of recombinant protein | |
| CN106632646A (en) | Method of co-purifing three microfunctional proteins from eggshell | |
| CN109852591B (en) | Method for extracting lipoxygenase from soybean whey wastewater by reverse pH gradient | |
| CN108003211A (en) | A kind of method of quick separating ganoderic acid in accessory substance from ganoderma lucidum | |
| US10301341B2 (en) | Technology for extracting and preparing high-purity raffinose from defatted wheat germ | |
| CN106589115B (en) | Method for separating and purifying soybean protease inhibitor | |
| CN106543266A (en) | A kind of method of scale purification recombination human apolipoprotein Apoa-I | |
| CN108727487B (en) | Liquid membrane extraction method of hirudin | |
| Ratanapariyanuch et al. | Industrial clarification of wheat-based distillers’ solubles and thin stillage | |
| CN103145592B (en) | Process for purifying special reagent sodium dodecyl sulfonate for ion pair chromatography | |
| Chandralekha et al. | Recent developments in the downstream processing of phycobiliproteins from algae: A review |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170510 |