CN105504023A - Capreomycin preparation method - Google Patents
Capreomycin preparation method Download PDFInfo
- Publication number
- CN105504023A CN105504023A CN201610097986.8A CN201610097986A CN105504023A CN 105504023 A CN105504023 A CN 105504023A CN 201610097986 A CN201610097986 A CN 201610097986A CN 105504023 A CN105504023 A CN 105504023A
- Authority
- CN
- China
- Prior art keywords
- preparation
- capromycin
- elutriant
- superparamagnetism microballoon
- capreomycin
- 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
- C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
- C07K7/50—Cyclic peptides containing at least one abnormal peptide link
- C07K7/54—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring
- C07K7/56—Cyclic peptides containing at least one abnormal peptide link with at least one abnormal peptide link in the ring the cyclisation not occurring through 2,4-diamino-butanoic acid
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Life Sciences & Earth Sciences (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a capreomycin preparation method. The capreomycin preparation method includes the steps of adding superparamagnetic microspheres into capreomycin fermentation liquor prior to adsorption, elution, decoloration, spray drying and the like so as to obtain a capreomycin product. The capreomycin preparation method has the advantages that pretreating and preheating the fermentation liquor during extraction are avoided, expensive extraction and separation equipment is unneeded, a production period is shortened by 2/3, pollution is reduced by 1/2, yield is increased by 10%, and accordingly technical support is provided for industrial production.
Description
Technical field
The invention belongs to industrial microbial technology field, be specifically related to a kind of preparation method of capromycin.
Background technology
Tuberculosis revives, positive serious threat human health.In recent years, the appearance of Drug-fast case tuberculosis (MDR-TB), and mycobacterium tuberculosis causes tuberculosis to become fashion trend in the whole world once again with the co-infection of human immunodeficiency virus.Capromycin is treatment substance of medicines-resistant branched tubercle bacillus and holds the class important drugs staying bacterium to infect.The pulmonary tuberculosis that this medicine causes drug-resistant type mycobacterium tuberculosis infection is evident in efficacy, and toxic side effect is little compared with other Second line Drugs, is current comparatively ideal anti-tubercle bacillus drugs.
Capromycin, be the ring type polypeptide class microbiotic produced by curling streptomycete fermentation, be made up of active compound IA, IB, IIA, IIB of four structural similitudies, wherein the content of IA and IB accounts for more than 90%, is the main component playing curative effect.
At present, the technical way of domestic production capreomycin sulfate, as patent CN100471867C, utilizes ion exchange resin to carry out adsorbing, desalination, decolouring, repeatedly to neutralize, form through concentrated, spraying dry again, process is complicated, is unfavorable for environmental protection, affects yield and quality product.
How can improve capromycin productive rate and quality, contriver has done lot of experiments.
Summary of the invention
The object of this invention is to provide the preparation method of the capromycin that a kind of technical process is short, easy to operate, product purity is high, environmental.
The preparation method of a kind of capromycin provided by the invention, comprises the following steps:
A, superparamagnetism microballoon is added in capromycin fermented liquid, stir and make it absorption completely, then isolate superparamagnetism microballoon.
B, isolated superparamagnetism microballoon is first used Water warfare, then use acidic aqueous solution wash-out, obtain capromycin elutriant.
C, add activated carbon decolorizing by capromycin elutriant, filter, obtain capromycin destainer.
D, capromycin destainer, through nanofiltration, spraying dry, obtain capromycin product.
Wherein the particle diameter of the described superparamagnetism microballoon of step (a) is 10-30 μm, and the functional group on surface is the sodium form structure after sodium hydroxide solution process.The add-on of described superparamagnetism microballoon adds 30-60 gram for often liter of fermented liquid.
The wherein aqueous sulfuric acid of the described sour water 0.2-0.5mol/L of step (b).Consumption is that every gram of superparamagnetism microballoon adds 4-6 milliliter.
Wherein the described activated carbon dosage of step (c) adds 0.5-1.0 gram for often liter of elutriant, bleaching time 30-40min.
The described nanofiltration of step (d) filtering membrane used is the rolled film of molecular weight cut-off 100-300Da.
The described spray-dired feed concentration of step (d) is 25-30 ten thousand μ g/mL.
The pre-treatment of superparamagnetism microballoon and regeneration, before use with the NaOH aqueous solution of the 2-5 doubly 2moL/L of (v/w), filter after static immersing 2-4 hour, then with deionized water wash to pH value most 7-8, for subsequent use.
Tool of the present invention has the following advantages: 1, adopt superparamagnetism microballoon, without the need to carrying out pre-treatment and preheating to fermented liquid in leaching process, can either the easy separation of realize target thing, the production cycle is for be reduced to 18h by 56h, efficiency improves 2/3, and discharging of waste liquid reduces 1/2; 2, superparamagnetism microballoon selectivity is high, and the extraction and isolation equipment without the need to costliness can obtain highly purified capromycin product, and quality product is better than standards of pharmacopoeia, and yield is greater than 85%, and the more former technique of yield improves 10%; 3, whole leaching process is not with an organic solvent, energy-conserving and environment-protective; 4, superparamagnetism microballoon have recyclable, recycle, regenerate the advantages such as easy; 5, the method for the invention technical process is short, overcomes the shortcoming that capromycin is difficult to purifying, and has easy to operate, that controllability is strong advantage, for suitability for industrialized production provides strong technical support.
Accompanying drawing explanation
Fig. 1 capromycin product high-efficient liquid phase chromatogram.
Embodiment
Further illustrate content of the present invention with specific embodiment below, but and mean never in any form and limit the invention.In the following example, method therefor is ordinary method if no special instructions.
Capromycin fermented liquid used in the present invention, for North China Pharmacuetical Group New Drug Research & Development Co., Ltd produces.Superparamagnetism microballoon, Suzhou Nano-Micro Bio-technology Co., Ltd.; Spray-drier (SP-1500), Shanghai is along instrument experimental installation company limited; High performance liquid chromatograph (SPD-M20A type UV-detector, LC-20AT pump, SIL-20A automatic sampler, CTO-10AS column oven, Shimadzu Corporation); Distilled water, for this laboratory is made by oneself; Other reagent are analytical pure.
The preparation of capromycin fermented liquid
The slant culture of curling streptomycete bacterial strain (numbering NRRL2773) or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 60h obtain seed liquor; Seed liquor is inoculated into fermentor tank and carries out fermentation culture, inoculum size is 10%, and culture temperature 29 DEG C, tank pressure 0.05 ± 0.01Mpa, ventilation ratio are that 1:0.8vvm, 400rpm stir, after fermentation culture starts 40 hours, often liter of fermented liquid adds 1.5g Methionin, and always ferment 144h.
Wherein said seed culture medium obtains by the following method: sucrose 30.0 grams, yeast powder 8.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 5.0 grams, 1.0 grams, sodium-chlor, 5.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
Wherein said fermention medium obtains by the following method: W-Gum 30.0 grams, cottonseed flour 20.0 grams, yeast powder 5.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 6.0 grams, potassium primary phosphate 2.0 grams, 2.0 grams, sodium-chlor, 5.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
The pre-treatment of superparamagnetism microballoon and regeneration
The pre-treatment of superparamagnetism microballoon and regeneration, before use with the NaOH aqueous solution of the 2-5 doubly 2moL/L of (v/w), filter after static immersing 2-4 hour, then with deionized water wash to pH value most 7-8, for subsequent use.
Embodiment 1
300mL fermented liquid, fermentation unit 11530 μ g/mL, add the superparamagnetism microballoon 18g that particle diameter is 10 μm, after whip attachment 1h, be separated superparamagnetism microballoon, with the isolated superparamagnetism microballoon of 36mL water washing, then the aqueous sulfuric acid wash-out superparamagnetism microballoon of 108mL0.2mol/L is used, collect capromycin elutriant, elutriant adds the activated carbon decolorizing of 0.054 gram, filter after stirring at room temperature 30min, filtrate again through the nanofiltration of 100Da rolled film to strength of solution 250,000 μ g/mL, carry out spraying dry, obtain capromycin product (high-efficient liquid phase chromatogram is shown in Fig. 1) 3.0g, content 98.6%, yield 85.5%.
Embodiment 2
1L fermented liquid, fermentation unit 11583 μ g/mL, add the superparamagnetism microballoon 50g that particle diameter is 20 μm, after whip attachment 1.5h, be separated superparamagnetism microballoon, with the isolated superparamagnetism microballoon of 100mL water washing, then the aqueous sulfuric acid wash-out superparamagnetism microballoon of 200mL0.3mol/L is used, collect capromycin elutriant, elutriant adds the gac of 0.16 gram, filter after stirring at room temperature 40min, filtrate again through the nanofiltration of 200Da rolled film to strength of solution 280,000 μ g/mL, carry out spraying dry, obtain capromycin product 10.1g, content 98.9%, yield 86.2%.
Embodiment 3
3L fermented liquid, fermentation unit 11516 μ g/mL, add the superparamagnetism microballoon 90g of particle diameter 30 μm, after whip attachment 2h, be separated superparamagnetism microballoon, with the isolated superparamagnetism microballoon of 180mL water washing, then use the aqueous sulfuric acid wash-out superparamagnetism microballoon of 450mL0.5mol/L, collect capromycin elutriant, elutriant adds the gac of 0.45 gram, filter after stirring 35min, filtrate again through the nanofiltration of 300Da rolled film to strength of solution 300,000 μ g/mL, carry out spraying dry, obtain capromycin product 30.3g, content 98.7%, yield 86.6%.
Comparative example 1
300mL fermented liquid, fermentation unit 11530 μ g/mL, after adding the dilution of 300mL water, adjust PH=3.0 with oxalic acid, then add 30g pearlite filtering aid, filter after stirring 30min, the NaOH solution of filtrate 2mol/L adjusts PH=7.0, then imports the ion exchange resin D116 that 100mL blade diameter length ratio is 1:10
-Naadsorb, adsorb complete, saturated resin is with after the purification of 300mL deionized water, and with the aqueous sulfuric acid wash-out of the 1mol/L of 300mL, it is 1:5 ion exchange resin 1 × 16 desalination that elutriant imports 30mL blade diameter length ratio, adds appropriate ion exchange resin 331 in demineralised liquid
-OH, stir and elutriant is neutralized to PH=6.0, neutralizer imports the ion exchange resin 122 that 50mL blade diameter length ratio is 1:10
-Hdecolouring, decolours complete, saturated resin use respectively 150mL0.5,1.0, the aqueous sulfuric acid gradient elution of 1.5mol/L, add appropriate ion exchange resin 331 in elutriant
-OH, stir and elutriant is neutralized to PH=7.0, namely obtain refined liquid, refined liquid is concentrated into 250,000 μ g/mL, carries out spraying dry, obtains capromycin product 2.8g, content 95.6%, yield 77.4%.
Comparative example 2
1L fermented liquid, fermentation unit 11583 μ g/mL, after adding the dilution of 1L water, PH=3.5 is adjusted with oxalic acid, then add after 160g flocculating aids stirs 30min and filter, the NaOH solution of filtrate 2mol/L adjusts PH=7.5, then imports the ion exchange resin D116 that 300mL blade diameter length ratio is 1:10
-Naadsorb, adsorb complete, saturated resin is with after the purification of 1000mL deionized water, and with the 2mol/L aqueous sulfuric acid wash-out of 700mL, it is 1:10 ion exchange resin 1 × 16 desalination that elutriant imports 100mL blade diameter length ratio, adds appropriate ion exchange resin 331 in demineralised liquid
-OH, stir and elutriant is neutralized to PH=7.0, neutralizer imports the ion exchange resin 122 that 150mL blade diameter length ratio is 1:10
-Hdecolouring, decolours complete, saturated resin with to use respectively 450mL0.5,1.0, the aqueous sulfuric acid gradient elution of 1.5mol/L, add appropriate ion exchange resin 331 in elutriant
-OH, stir and elutriant is neutralized to PH=6.5, namely obtain refined liquid, refined liquid is concentrated into 300,000 μ g/mL, carries out spraying dry, obtains capromycin product 9.2g, content 96.2%, yield 76.4%.
Comparative example 3
3L fermented liquid, fermentation unit 11516 μ g/mL, after adding the dilution of 3L water, PH=3.2 is adjusted with oxalic acid, then add after 600g flocculating aids stirs 30min and filter, the NaOH solution of filtrate 2mol/L adjusts PH=8.0, then imports the ion exchange resin D116 that 1L blade diameter length ratio is 1:10
-Naadsorb, adsorb complete, saturated resin is with after the purification of 3L deionized water, and with the 1.5mol/L aqueous sulfuric acid wash-out of 2.5L, it is 1:10 ion exchange resin 1 × 16 desalination that elutriant imports 300mL blade diameter length ratio, adds appropriate ion exchange resin 331 in demineralised liquid
-OH, stir and elutriant is neutralized to PH=6.5, neutralizer imports the ion exchange resin 122 that 500mL blade diameter length ratio is 1:10
-Hdecolouring, decolours complete, saturated resin with to use respectively 1500mL0.5,1.0, the aqueous sulfuric acid gradient elution of 1.5mol/L, add appropriate ion exchange resin 331 in elutriant
-OH, stir and elutriant is neutralized to PH=7.0, namely obtain refined liquid, refined liquid is concentrated into 300,000 μ g/mL, carries out spraying dry, obtains capromycin product 27.7g, content 95.8%, yield 76.8%.
Claims (8)
1. a preparation method for capromycin, comprises the steps:
A, superparamagnetism microballoon is added in capromycin fermented liquid, stir and make it absorption completely, then isolate superparamagnetism microballoon;
B, isolated superparamagnetism microballoon is first used Water warfare, then use acidic aqueous solution wash-out, obtain capromycin elutriant;
C, add activated carbon decolorizing by capromycin elutriant, filter, obtain capromycin destainer;
D, capromycin destainer, through nanofiltration, spraying dry, obtain capromycin product.
2. preparation method according to claim 1, wherein the particle diameter of the described superparamagnetism microballoon of step (a) is 10-30 μm, and the functional group on surface is the sodium form structure after sodium hydroxide solution process.
3. preparation method according to claim 1, wherein the add-on of the described superparamagnetism microballoon of step (a) adds 30-60 gram for often liter of fermented liquid.
4. preparation method according to claim 1, wherein the described sour water of step (b) is the aqueous sulfuric acid of 0.2-0.5mol/L.
5. preparation method according to claim 1, wherein the described sour water consumption of step (b) is that every gram of superparamagnetism microballoon adds 4-6 milliliter.
6. preparation method according to claim 1, wherein the described activated carbon dosage of step (c) adds 0.5-1.0 gram for often liter of elutriant.
7. preparation method according to claim 1, the filtering membrane that wherein the described nanofiltration of step (d) is used is the rolled film of molecular weight cut-off 100-300Da.
8. preparation method according to claim 1, wherein the described spray-dired feed concentration of step (d) is 25-30 ten thousand μ g/mL.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610097986.8A CN105504023A (en) | 2016-02-23 | 2016-02-23 | Capreomycin preparation method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610097986.8A CN105504023A (en) | 2016-02-23 | 2016-02-23 | Capreomycin preparation method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105504023A true CN105504023A (en) | 2016-04-20 |
Family
ID=55712376
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610097986.8A Pending CN105504023A (en) | 2016-02-23 | 2016-02-23 | Capreomycin preparation method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105504023A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100471867C (en) * | 2003-06-18 | 2009-03-25 | 南宁中科药业有限责任公司 | Novel method for abstracting vitriol capreomycin |
| CN103065754A (en) * | 2012-12-04 | 2013-04-24 | 天津大学 | Magnetic material for surface finish benzene sulfonic acid, and preparation method and application thereof |
| CN103076415A (en) * | 2011-11-15 | 2013-05-01 | 南昌大学 | Method for enriching nature tetracycline antibiotics in aquatic products |
| CN105131091A (en) * | 2015-07-30 | 2015-12-09 | 宁夏泰瑞制药股份有限公司 | Method for preparing capreomycin sulfate from capreomycin broth |
-
2016
- 2016-02-23 CN CN201610097986.8A patent/CN105504023A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100471867C (en) * | 2003-06-18 | 2009-03-25 | 南宁中科药业有限责任公司 | Novel method for abstracting vitriol capreomycin |
| CN103076415A (en) * | 2011-11-15 | 2013-05-01 | 南昌大学 | Method for enriching nature tetracycline antibiotics in aquatic products |
| CN103065754A (en) * | 2012-12-04 | 2013-04-24 | 天津大学 | Magnetic material for surface finish benzene sulfonic acid, and preparation method and application thereof |
| CN105131091A (en) * | 2015-07-30 | 2015-12-09 | 宁夏泰瑞制药股份有限公司 | Method for preparing capreomycin sulfate from capreomycin broth |
Non-Patent Citations (3)
| Title |
|---|
| 张天兵: "膜技术在卷曲霉素提炼工艺中的推广应用", 《中国学位论文全文数据库》 * |
| 许学受 等: "卷曲霉素的综合介绍", 《安徽医学》 * |
| 邓小娟: "基于磁性纳米颗粒的功能化复合材料的制备及其吸附性能研究", 《中国博士学位论文全文数据库（电子期刊）工程科技I辑》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105294910A (en) | Preparation method of sorbent applied to pristinamycin separation | |
| CN102936253B (en) | A kind of preparation method of high purity tacrolimus | |
| CN102718843B (en) | Preparation method of single teicoplanin components | |
| CN103304640B (en) | A kind of method extracting Echinocandin compound from fermented liquid | |
| CN103275152B (en) | A kind of preparation method of high-purity fidaxomicin | |
| CN105481950A (en) | Daptomycin extraction method | |
| CN109161684A (en) | A method of utilizing spirulina recovering rare earth | |
| CN107792911A (en) | A kind of method for going in water removal to remain tetracycline using absorption method | |
| CN105566458A (en) | Method for preparing bacitracin and method for preparing zinc salt of bacitracin | |
| CN104289199A (en) | Preparation method of magnetic HPCS/GO adsorbing agent for removing heavy metals and application | |
| CN103224547B (en) | Daptomycin separation and purification method | |
| CN105504023A (en) | Capreomycin preparation method | |
| CN101838315B (en) | Method for separating Ramoplanin | |
| CN112409426B (en) | Preparation method of sisomicin sulfate | |
| CN103073624B (en) | A kind of preparation method of high purity cyclosporin A derivative | |
| CN103276028B (en) | A kind of method that fermentation method produces epothilone B | |
| CN105441504A (en) | Preparation method of cyclic dipeptide compound | |
| CN102127127A (en) | Kanamycin extracting process | |
| CN114057753B (en) | Method for separating and purifying antifungal active substance HSAF in zymogen fermentation liquor | |
| CN110590883B (en) | Method for extracting and separating spinosad from saccharopolyspora spinosa fermentation liquor by adopting resin | |
| CN106032547B (en) | Preparation method and purification method of ergosterol compound | |
| CN103012515B (en) | Preparation method of high-purity gentamycin | |
| CN101602785B (en) | Method for quickly desalting and purifying selenizing oligosaccharide | |
| CN107417750B (en) | Method for extracting cyclic adenosine monophosphate from microbial fermentation liquid | |
| CN109735713A (en) | A method of utilizing metal-organic framework material UiO-66 adsorbing separation indium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into 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: 20160420 |