CN103232525B - Method for extracting polyoxin from streptomyces fermentation broth - Google Patents
Method for extracting polyoxin from streptomyces fermentation broth Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 22
- 238000000855 fermentation Methods 0.000 title claims abstract description 15
- 230000004151 fermentation Effects 0.000 title claims abstract description 15
- 229930182764 Polyoxin Natural products 0.000 title abstract description 6
- YEBIHIICWDDQOL-YBHNRIQQSA-N polyoxin Polymers O[C@@H]1[C@H](O)[C@@H](C(C=O)N)O[C@H]1N1C(=O)NC(=O)C(C(O)=O)=C1 YEBIHIICWDDQOL-YBHNRIQQSA-N 0.000 title abstract description 5
- 241000187747 Streptomyces Species 0.000 title abstract 3
- 238000001953 recrystallisation Methods 0.000 claims abstract description 9
- 238000001291 vacuum drying Methods 0.000 claims abstract description 5
- 239000007788 liquid Substances 0.000 claims description 39
- 238000010025 steaming Methods 0.000 claims description 29
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- 230000003115 biocidal effect Effects 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 15
- 239000013078 crystal Substances 0.000 claims description 12
- 239000003960 organic solvent Substances 0.000 claims description 11
- 239000002253 acid Substances 0.000 claims description 9
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 229920005654 Sephadex Polymers 0.000 claims description 7
- 239000012507 Sephadex™ Substances 0.000 claims description 7
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical group CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- 241001655322 Streptomycetales Species 0.000 claims description 6
- 150000003839 salts Chemical class 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 229960000935 dehydrated alcohol Drugs 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 2
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 2
- 239000012498 ultrapure water Substances 0.000 claims description 2
- 238000000926 separation method Methods 0.000 abstract description 3
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 abstract 1
- 239000003729 cation exchange resin Substances 0.000 abstract 1
- 238000011033 desalting Methods 0.000 abstract 1
- 238000002845 discoloration Methods 0.000 abstract 1
- 238000010828 elution Methods 0.000 abstract 1
- 238000005516 engineering process Methods 0.000 abstract 1
- 238000001914 filtration Methods 0.000 abstract 1
- 239000011347 resin Substances 0.000 abstract 1
- 229920005989 resin Polymers 0.000 abstract 1
- 238000001179 sorption measurement Methods 0.000 abstract 1
- 229920002101 Chitin Polymers 0.000 description 7
- WURKLEUBADSHTC-UHFFFAOYSA-N [6-[6-[6-[[6-[4-acetyloxy-5-(4-hydroxy-5-methoxy-6-methyloxan-2-yl)oxy-6-methyloxan-2-yl]oxy-3-(3,4-dihydroxy-1-methoxy-2-oxopentyl)-8,9-dihydroxy-1-oxo-3,4-dihydro-2h-anthracen-2-yl]oxy]-3-hydroxy-2-methyloxan-4-yl]oxy-3-hydroxy-2-methyloxan-4-yl]oxy-4-h Chemical compound C=1C(O)=C2C(O)=C3C(=O)C(OC4OC(C)C(O)C(OC5OC(C)C(O)C(OC6OC(C)C(OC(=O)C(C)C)C(C)(O)C6)C5)C4)C(C(OC)C(=O)C(O)C(C)O)CC3=CC2=CC=1OC(OC1C)CC(OC(C)=O)C1OC1CC(O)C(OC)C(C)O1 WURKLEUBADSHTC-UHFFFAOYSA-N 0.000 description 4
- 102000003960 Ligases Human genes 0.000 description 3
- 108090000364 Ligases Proteins 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 239000006067 antibiotic powder Substances 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- -1 phosphorus-N-Acetyl-D-glucosamine structure Chemical group 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 241000221785 Erysiphales Species 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical class CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 1
- 241000896238 Oidium Species 0.000 description 1
- 208000031320 Teratogenesis Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229960000074 biopharmaceutical Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000315 carcinogenic Toxicity 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000002860 competitive effect Effects 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- HWJHWSBFPPPIPD-UHFFFAOYSA-N ethoxyethane;propan-2-one Chemical compound CC(C)=O.CCOCC HWJHWSBFPPPIPD-UHFFFAOYSA-N 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 230000002070 germicidal effect Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000010534 mechanism of action Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000002718 pyrimidine nucleoside Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
Abstract
The invention discloses a method for extracting polyoxin from a streptomyces fermentation broth, and belongs to the field of biological engineering. According to the invention, through streptomyces fermentation broth filtering, discoloration, cation exchange resin adsorption, elution, gel resin desalting, re-crystallization, and vacuum drying technologies, the polyoxin is obtained. The method is a safe and reliable separation method with the advantages of easy operation and low cost.
Description
Technical field
The present invention relates to a kind of technique of many antibiotic of purifying from fermented liquid, belong to bioengineering field, be specifically related to bio-pharmaceuticals, the extraction of tunning, the method for separation, particularly a kind of method proposing many antibiotic from streptomycete fermentation liquid.
Technical background
Many antibiotic (Polyoxins) is also referred to as multi-effect mycin, polyoxin, Polyoxin, is a kind of peptide pyrimidine nucleoside acids agricultural antibiotic.Research finds, 14 components (polyoxinA-N) that many antibiotic is correlated with by structure form.The fusing point that they do not fix, decomposes gradually at about 200 DEG C, and soluble in water, is insoluble in the organic solvents such as methyl alcohol, ethanol, butanols, acetone ether, comparatively stable in the scope that pH is 1-8.
Many antibiotic, as a kind of broad-spectrum antibiotics, has the target organisms selectivity of good interior absorption and height.Many antibiotic mechanism of action suppresses the katalysis of chitin synthetase, causes the composition chitin of fungal cell wall not synthesize and to play germicidal action.It is similar that the substrate of many antibiotic and chitin synthetase urinates two phosphorus-N-Acetyl-D-glucosamine structure, thus suppress by chitin synthesis enzyme catalysis, by urine two phosphorus-N-Acetyl-D-glucosamine to this single step reaction of chitin, by the dynamics research of enzyme, prove that many antibiotic is the competitive inhibitor of chitin synthetase.It has excellent effect to multiple fungal diseases such as preventing and treating gray mold, oidium, early blight, bloom disease, Powdery Mildew, blight.Due in general farm crop and mammalian body without chitin integral part, therefore many antibiotic is to person poultry safety, without summation, without " three-induced effect " (mutagenesis, teratogenesis, carcinogenic).
For a long time, many antibiotic is monopolized by Japanese Kaken Pharmaceutical Co., Ltd, also has manufacturer production in the past although Chinese, but be no matter on quality product or fermentation unit, all cannot contend with Japan, export volume is little, is be limited on the cash crop of domestic market to apply substantially.Because Japanese like product selling price is high, cause drug cost relatively high, mainly as the high-end sterilant of one in application, thus hinder the large-scale promotion application of this kind.At present the most research of many antibiotic is mainly concentrated on to fermentation and the context of detection of many antibiotic, the report that aobvious rare many antibiotic is separated.Therefore, this area in the urgent need to providing a kind of easy handling, cost low, safety, reliable separation method.
Summary of the invention
The object of the invention is to provide a kind of method extracting many antibiotic from streptomycete fermentation liquid to improve the deficiencies in the prior art, adopt the method for ion exchange technique and recrystallization from fermented liquid, extract the technique of many antibiotic, easy to operate, technique simple, reduce costs, raise the efficiency.
Technical scheme of the present invention is: a kind of method extracting many antibiotic from streptomycete fermentation liquid, and its concrete steps are as follows:
(1) streptomycete fermentation liquid is got, with acid for adjusting pH to 1.0-4.0, fermented liquid is centrifugal, collect supernatant liquor, filter, obtain clarified liq;
(2) clarified liq of step (1) gained is poured in separating funnel, then add organic solvent wherein, vibration, leave standstill 0.5-2h, get supernatant liquor, revolve steaming, steaming liquid must be revolved;
(3) revolving of step (2) gained is steamed liquid acid for adjusting pH to 1.0-4.0, transfer to Zeo-karb is housed bed on adsorb, and carrying out wash-out with salts solution, every 2-5min collects an elutriant, is eluted to till elutriant ultraviolet spectrophotometer do not detect peak; Be associated with the elutriant detecting peak, carry out revolving steaming, obtain concentrated solution;
(4) concentrated solution of step (3) gained is transferred to sephadex column, carries out wash-out with elutriant, every 5-10min collects once, is eluted to till elutriant ultraviolet spectrophotometer do not detect peak, is associated with the elutriant detecting peak;
(5) elutriant of step (4) gained is revolved steaming, concentrated;
(6) by step (5) gained concentrated solution acid for adjusting pH value to 1.0-4.0, then add organic solvent, until crystal is no longer separated out, obtain recrystallization crystal;
(7) the recrystallization crystal vacuum drying oven in step (6) is carried out drying.
Rotating speed centrifugal in preferred steps (1) is 8000-10000rmp/min; Centrifugation time is 15-25min.Preferred steps (1), (3) and the acid described in (6) are HCl, H
2sO
4or H
3pO
4solution, its concentration is 2.0-6.0mol/L.
Organic solvent described in preferred steps (2) is ethyl acetate, and the volume ratio of organic solvent and clarified liq is 0.1-0.5:1; After revolving steaming, revolving the volume ratio of steaming liquid and supernatant liquor is 1:(50-100).
The kind of Zeo-karb described in preferred steps (3) is 001 × 4,001 × 7 or 110 type Zeo-karbs, and ion column footpath and height are than being 1:(8-12); Salts solution is NaCl or KCl solution, and its mass percentage concentration is 10-30%, and the flow velocity of salts solution is 5-15mL/min; Revolving the volume ratio of steaming its volume of liquid and elutriant is 1:(50-100).
Preferred steps (3) and (4) medium ultraviolet spectrophotometric many antibiotic detected peaks scope are between 240-280nm.
Sephadex column described in preferred steps (4) is G-15 or G-25, and gel column footpath and height are 1:(5-7); Elutriant is deionized water or ultrapure water; The flow velocity of elutriant is 0.5-2mL/min.
Revolving described in preferred steps (5) steams the volume of liquid and the volume ratio of elutriant is 1:(50-100).
Organic solvent described in preferred steps (6) is dehydrated alcohol or acetone.
The temperature of the vacuum drying oven described in preferred steps (7) is 60 ~ 80 DEG C, and vacuum tightness is-0.075 ~-0.01MPa, and time of drying is 12 ~ 24 hours.
Embodiment
Explain the present invention further below in conjunction with example, but case study on implementation does not limit in any form to the present invention.
Embodiment 1
(1) get streptomycete fermentation liquid, regulate pH to 1.0 with the HCl solution that concentration is 2.0mol/L, by fermented liquid with the centrifugal 15min of the rotating speed of 8000rmp/min, collect supernatant liquor, use filtered on buchner funnel, obtain clarified liq;
(2) get the clear liquor of step (1) gained, pour in separating funnel, then add ethyl acetate wherein, the volume ratio of ethyl acetate and clear liquor is 0.1:1, vibration, leaves standstill 0.5h, gets supernatant liquor, revolve steaming, revolves and steams liquid with supernatant volume than being 1:50.
(3) revolve that to steam liquid concentration be the H of 3.0mol/L by step (2) gained
2sO
4solution regulates pH to 3.0, transfer to 001 × 7 type Zeo-karb is housed bed on adsorb, ion column footpath/height compares 1:8, and with mass percent be 10% NaCl solution carry out wash-out with the speed of 5mL/min, elutriant is collected with Fraction Collector, every 2min collects once, is eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 240-270nm.Be incorporated within the scope of 240-270nm the elutriant having and detect peak, carry out revolving steaming, revolving the volume ratio of steaming liquid and elutriant is 1:70, obtains concentrated solution;
(4) concentrated solution of step (3) gained is transferred to sephadex column G-25, gel column footpath/height compares 1:5, wash-out is carried out with the speed of 0.5mL/min with deionized water, every 5min collects once, be eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 240-270nm, be incorporated within the scope of 240-270nm the elutriant having and detect peak;
(5) elutriant of step (4) gained is revolved steaming, revolving the volume ratio of steaming liquid and elutriant is 1:50;
(6) to step (5) gained concentrated solution concentration be the HCl solution adjust ph to 4.0 of 6.0mol/L, then add dehydrated alcohol wherein, obtain recrystallization crystal;
(7) crystal in step (6) is put into vacuum tightness loft drier, temperature sets 60 DEG C, and vacuum tightness is-0.075MPa, dry 12 hours, obtains powder, detects, obtain many antibiotic powder through HPLC; Productive rate is 81%, and purity is 98.7%.
Embodiment 2
(1) getting streptomycete fermentation liquid, is the H of 6.0mol/L by concentration
2sO
4solution regulates pH to 4.0, by fermented liquid with the centrifugal 25min of the rotating speed of 10000rmp/min, collects supernatant liquor, uses filtered on buchner funnel, obtain clarified liq;
(2) get the clarified liq of step (1) gained, pour in separating funnel, then add ethyl acetate wherein, the volume ratio of ethyl acetate and clear liquor is 0.5:1, vibration, leaves standstill 2h, gets supernatant liquor, revolve steaming, revolves and steams liquid with supernatant volume than being 1:100.
(3) revolving of step (2) gained is steamed the HCl solution adjustment pH to 1.0 that liquid concentration is 4.0mol/L, transfer to 001 × 4 type Zeo-karb is housed bed on adsorb, ion column footpath/height compares 1:12, and carry out wash-out with the KCl solution that mass percent is 30% with the speed of 15mL/min, elutriant is collected with Fraction Collector, every 5min collects once, is eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 250-280nm.Be incorporated within the scope of 250-280nm the elutriant having and detect peak, carry out revolving steaming, revolving the volume ratio of steaming liquid and elutriant is 1:50, obtains concentrated solution;
(4) concentrated solution of step (3) gained is transferred to sephadex column G-15, gel column footpath/height compares 1:7, wash-out is carried out with the speed of 2mL/min with deionized water, every 10min collects once, be eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 250-280nm, be incorporated within the scope of 250-280nm the elutriant having and detect peak;
(5) elutriant of step (4) gained is revolved steaming, revolving the volume ratio of steaming liquid and elutriant is 1:100;
(6) to step (5) gained concentrated solution concentration be the H of 2.0mol/L
2sO
4solution adjust ph to 3.0, more slowly add dehydrated alcohol wherein, obtain recrystallization crystal;
(7) crystal in step (6) is put into vacuum tightness loft drier, temperature sets 80 DEG C, and vacuum tightness is-0.01MPa, dry 24 hours, obtains powder, detects, obtain many antibiotic powder through HPLC; Productive rate is 87%, and purity is 99.2%.
Embodiment 3
(1) get streptomycete fermentation liquid, regulate pH to 3.0 with the HCl solution that concentration is 5.0mol/L, by fermented liquid with the centrifugal 18min of the rotating speed of 9000rmp/min, collect supernatant liquor, use filtered on buchner funnel, obtain clarified liq;
(2) get the clarified liq of step (1) gained, pour in separating funnel, then add ethyl acetate wherein, the volume ratio of ethyl acetate and clear liquor is 0.25:1, vibration, leaves standstill 1.5h, gets supernatant liquor, revolve steaming, revolves and steams liquid with supernatant volume than being 1:100.
(3) revolve that to steam liquid concentration be the H of 2.0mol/L by step (2) gained
2sO
4solution regulates pH to 2.0, transfer to 110 type Zeo-karbs are housed bed on adsorb, ion column footpath/height compares 1:10, and with mass percent be 30% NaCl solution carry out wash-out with the speed of 10mL/min, elutriant is collected with Fraction Collector, every 5min collects once, is eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 240-280nm.Be incorporated within the scope of 240-280nm the elutriant having and detect peak, carry out revolving steaming, revolving the volume ratio of steaming liquid and elutriant is 1:100, obtains concentrated solution;
(4) concentrated solution of step (3) gained is transferred to sephadex column G-25, gel column footpath/height compares 1:6, wash-out is carried out with the speed of 2mL/min with deionized water, every 10min collects once, be eluted to elutriant UV spectrophotometer measuring, until do not detect peak within the scope of 250-280nm, be incorporated within the scope of 250-280nm the elutriant having and detect peak;
(5) elutriant of step (4) gained is revolved steaming, revolving the volume ratio of steaming liquid and elutriant is 1:70;
(6) to step (5) gained concentrated solution concentration be the HCl solution adjust ph to 1.0 of 3.0mol/L, more slowly add acetone wherein, obtain recrystallization crystal;
(7) crystal in step (6) is put into vacuum tightness loft drier, temperature sets 60 DEG C, and vacuum tightness is-0.070MPa, dry 20 hours, obtains powder, detects, obtain many antibiotic powder through HPLC; Productive rate is 83%, and purity is 98.3%.
Claims (9)
1. from streptomycete fermentation liquid, extract a method for many antibiotic, its concrete steps are as follows:
(1) streptomycete fermentation liquid is got, with acid for adjusting pH to 1.0-4.0, fermented liquid is centrifugal, collect supernatant liquor, filter, obtain clarified liq;
(2) clarified liq of step (1) gained is poured in separating funnel, then add organic solvent wherein, vibration, leave standstill 0.5-2h, get supernatant liquor, revolve steaming, steaming liquid must be revolved;
(3) revolving of step (2) gained is steamed liquid acid for adjusting pH to 1.0-4.0, transfer to Zeo-karb is housed bed on adsorb, and carry out wash-out with salts solution, every 2-5min collects an elutriant, is eluted to till elutriant ultraviolet spectrophotometer do not detect peak; Be associated with the elutriant detecting peak, carry out revolving steaming, obtain concentrated solution; The kind of wherein said Zeo-karb is 001 × 4,001 × 7 or 110 type Zeo-karbs, and ion column footpath and height are than being 1:(8-12);
(4) concentrated solution of step (3) gained is transferred to sephadex column, carries out wash-out with elutriant, every 5-10min collects once, is eluted to till elutriant ultraviolet spectrophotometer do not detect peak, is associated with the elutriant detecting peak; Wherein said sephadex column is G-15 or G-25, and gel column footpath and height are 1:(5-7);
(5) elutriant of step (4) gained is revolved steaming, concentrated;
(6) by step (5) gained concentrated solution acid for adjusting pH value to 1.0-4.0, then add organic solvent, until crystal is no longer separated out, obtain recrystallization crystal; Described organic solvent is dehydrated alcohol or acetone;
(7) the recrystallization crystal vacuum drying oven in step (6) is carried out drying.
2. method according to claim 1, is characterized in that: rotating speed centrifugal in step (1) is 8000-10000rpm; Centrifugation time is 15-25min.
3. method according to claim 1, is characterized in that: step (1), (3) and the acid described in (6) are HCl, H
2sO
4or H
3pO
4solution, its concentration is 2.0-6.0mol/L.
4. method according to claim 1, is characterized in that: the organic solvent described in step (2) is ethyl acetate, and the volume ratio of organic solvent and clarified liq is 0.1-0.5:1; After revolving steaming, revolving the volume ratio of steaming liquid and supernatant liquor is 1:(50-100).
5. method according to claim 1, is characterized in that: the salts solution in step (3) is NaCl or KCl solution, and its mass percentage concentration is 10-30%, and the flow velocity of salts solution is 5-15mL/min; Revolving the volume ratio of steaming its volume of liquid and elutriant is 1:(50-100).
6. method according to claim 1, is characterized in that: step (3) and (4) medium ultraviolet spectrophotometric many antibiotic detected peaks scope are between 240-280nm.
7. method according to claim 1, is characterized in that: the elutriant in step (4) is deionized water or ultrapure water; The flow velocity of elutriant is 0.5-2mL/min.
8. method according to claim 1, is characterized in that: revolving described in step (5) steams the volume of liquid and the volume ratio of elutriant is 1:(50-100).
9. method according to claim 1, is characterized in that: the temperature of the vacuum drying oven described in step (7) is 60 ~ 80 DEG C, and vacuum tightness is-0.075 ~-0.01MPa, and time of drying is 12 ~ 24 hours.
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| CN103613641B (en) * | 2013-11-11 | 2015-07-08 | 南京工业大学 | Method for separating and purifying polyoxins |
| CN106635878A (en) * | 2016-10-25 | 2017-05-10 | 扬州大学 | Streptomyces sp. 5X4 and applications thereof as biocontrol bacterium in preventing and treating plant diseases |
| CN109781886A (en) * | 2019-01-30 | 2019-05-21 | 陕西麦可罗生物科技有限公司 | A kind of preparation method and detection method of polyoxin N component |
| CN109765315A (en) * | 2019-01-30 | 2019-05-17 | 陕西麦可罗生物科技有限公司 | A kind of preparation method and detection method of polyoxin component C |
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| CN1436787A (en) * | 2002-02-04 | 2003-08-20 | 中国农业科学院植物保护研究所 | Antibiotic and its prepn and application |
| CN102153633A (en) * | 2010-12-27 | 2011-08-17 | 江苏九阳生物制药有限公司 | Method for extracting and separating bacitracin |
| CN102532277A (en) * | 2011-12-26 | 2012-07-04 | 中国科学院微生物研究所 | Agricultural antibiotic polyoxin P, biosynthesis method and application thereof |
| CN102603867A (en) * | 2012-02-29 | 2012-07-25 | 武汉大学 | Polyoxin-nikkomycin hybrid antibiotic and preparation method thereof |
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