CN105506042A - Method for producing capreomycin by fermentation - Google Patents

Method for producing capreomycin by fermentation Download PDF

Info

Publication number
CN105506042A
CN105506042A CN201610097987.2A CN201610097987A CN105506042A CN 105506042 A CN105506042 A CN 105506042A CN 201610097987 A CN201610097987 A CN 201610097987A CN 105506042 A CN105506042 A CN 105506042A
Authority
CN
China
Prior art keywords
fermentation
gram
grams
capreomycin
capromycin
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
Application number
CN201610097987.2A
Other languages
Chinese (zh)
Inventor
高任龙
王耀耀
张雪霞
任风芝
耿文飞
路新华
段宝玲
杨海静
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NCPC New Drug Research and Development Co Ltd
Original Assignee
NCPC New Drug Research and Development Co Ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by NCPC New Drug Research and Development Co Ltd filed Critical NCPC New Drug Research and Development Co Ltd
Priority to CN201610097987.2A priority Critical patent/CN105506042A/en
Publication of CN105506042A publication Critical patent/CN105506042A/en
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/465—Streptomyces

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • General Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention belongs to the field of fermentation engineering and particularly relates to a method for producing capreomycin by fermentation. The method ensures stability of inter-batch fermentation units by optimizing a culture medium and fermentation conditions; by adding precursor material in a fermentation process, production of capreomycin can be significantly promoted; by adding super-paramagnetic microspheres in a capreomycin synthesis phase to adsorb capreomycin in a fermentation broth, feedback inhibition of a fermentation product upon generated bacteria is lowered. After fermentation, fermentation unit of the capreomycin is up to 14149 ug/ml; compared with pre-optimization methods, this method provides an increase of 73% in capreomycin yield, an improvement in productivity and a reduction in energy consumption and waste discharge.

Description

A kind of method of fermentative production capromycin
Technical field
The invention belongs to field of fermentation engineering, be specifically related to a kind of method of fermentative production 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.The molecular formula of IA is C 25h 44n 14o8, molecular weight is the molecular formula of 668, IB is C 25h 44n 14o 7, molecular weight is 652.
In capromycin fermenting process, add precursor substance is the important means improving fermentation unit, by adding precursor substance, can reach the biosynthesizing direction controlling curling streptomycete and the object increasing capromycin output.Capromycin is ring type polypeptide class microbiotic, follows NRPS(NRPS114) biosynthetic pathway, its precursor substance is specific amino acid.
In the biosynthetic pathway of capromycin, mainly comprise the synthesis of non-protein amino acid, the biosynthesizing of peptide ring skeleton, Cheng Huanhou structural modification.The peptide ring skeleton of capromycin is the 5 peptide rings catalyzed and synthesized through Nonribosomal Peptide Synthetases by multiple non-protein amino acid through modification and natural amino acid.β carbamoylation on Cheng Huanhou amino-acid residue 4; β Methionin acylations on amino-acid residue 3 becomes important feature modification step; under the effect of amino acid activating cnzyme and peptide condensing enzyme, form side ingredient intermediate Capreomycin II A, IIB conversion to active principle IA, IB.Add the amino acid precursor material of synthesis capromycin from the external world, the fermentation level of capromycin can be improved significantly.But the residual concentration of exogenous precursor substance in fermented liquid is too high, production bacterial strain can be made poisoning, be unfavorable for the biosynthesizing of capromycin; And precursor substance deficiency also can affect the production of capromycin greatly.Therefore, study suitable precursor interpolation strategy to have great importance to the stable high yield of capromycin.
Capromycin is the secondary metabolite of fermentable, is present in fermented liquid, and capromycin not only kills or restraining effect other biological, also kills or restraining effect producing strains self, and namely microbiotic also exists toxic action to producing strains self.Along with capromycin concentration in fermented liquid constantly increases, its producing strains is subject to the feedback regulation effect of high density product, the potentiality of producing strains biomass cells and enzyme can not be given full play to, cause the productive rate of capromycin to be difficult to improve, thus become the bottleneck that restriction improves capromycin fermentation level.The producing strains of seed selection energy enduring high-concentration capromycin no doubt can make productive capacity be improved, but can not tackle the problem at its root.
Summary of the invention
The object of the invention is to solve fermentable in prior art and prepare the low problem of fermentation level that capromycin exists, there is provided a kind of by optimizing fermentative medium formula, fermentation condition, add precursor substance during the fermentation, promote the biosynthesizing of capromycin, thus improve the method for capromycin output.
The method of a kind of fermentative production capromycin provided by the present invention, comprises the following steps:
A, prepare the seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 26-30 DEG C, cultivation 48-72h acquisition seed liquor.
Wherein said seed culture medium obtains by the following method: sucrose 20.0-40.0 gram, yeast powder 5.0-10.0 gram, ammonium sulfate 2.0-3.0 gram, corn steep liquor 2.0-8.0 gram, sodium-chlor 0.5-2.0 gram, calcium carbonate 3.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention can select the bacterial classification of bacterium numbering NRRL2773.
B, prepare the fermented liquid of curling streptomycete bacterial strain
Above-mentioned seed liquor is inoculated in fermention medium with the inoculum size of volume percent 7-10%, in fermentation cylinder for fermentation, ferments between 24-56 hour and add precursor substance Methionin; The total fermentation time 120-144 time, culture temperature 27-29 DEG C, obtains fermented liquid.
Wherein said fermention medium obtains by the following method: W-Gum 20.0-30.0 gram, cottonseed flour 0-20.0 gram, soybean cake powder 0-20.0 gram, yeast powder 0-5.0 gram, ammonium sulfate 2.0-4.0 gram, corn steep liquor 4.0-8.0 gram, potassium primary phosphate 2.0-4.0 gram, sodium-chlor 1.0-2.0 gram, calcium carbonate 4.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
Further improvement of the present invention is: in described step b, precursor substance addition adds 0.5-2g for often liter of fermented liquid.
Further improvement of the present invention is: the tank pressure that ferments in described step b is 0.05 ± 0.01Mpa, ventilation ratio is 1:0.8-1.2vvm, stirring velocity is 300-400rpm.
Further improvement of the present invention is: ferment in described step b between 48-72 hour and add the superparamagnetism microballoon after sterilizing.
Further improvement of the present invention is: described superparamagnetism microspherulite diameter is 5-30um, and size distribution is single dispersing, and the functional group on surface is hydroxy-acid group, and superparamagnetism microballoon is through the sodium form structure after sodium hydroxide solution process.
Further improvement of the present invention is: superparamagnetism microballoon addition adds 10-20g for often liter of fermented liquid.
Described superparamagnetism microballoon with the NaOH aqueous solution of the 2-5 doubly 2moL/L of (v/w), filters after static immersing 2-4 hour before use, then with deionized water wash to pH value most 7-8, be placed in encloses container, 120 DEG C, pressure is under 97kPa, sterilizing 30min.After having fermented, the capromycin of acidic aqueous solution wash-out absorption used by superparamagnetism microballoon after externally-applied magnetic field reclaims.
Owing to have employed technique scheme, the technical progress that the present invention obtains is:
Curling streptomycete bacterial strain is adopted to be starting strain, produce in the process of amphotericin B at fermentable, optimize the conditions such as initial pH value, leavening temperature and time, inoculum size, substratum, in the capromycin Product formation phase, add appropriate precursor substance, while promotion effectively synthesis capromycin, turn avoid bacterial strain poisoning.Also by adding superparamagnetism microballoon, reducing product to the feedback inhibition of producing strains, thus significantly improving capromycin output.
Beneficial effect of the present invention:
Above-mentioned optimum condition, obtain capromycin output bring up to 14149ug/ml from 8166ug/ml, the suitability for industrialized production for capromycin provides a kind of method.By optimization of C/C composites and fermentation condition in the present invention, ensure that a batch stability for indirect fermentation unit; Significantly can promote the biosynthesizing of capromycin after adding precursor substance, and not add compared with precursor substance, fermentation unit improves more than 20%; Add the capromycin in adsorbable fermented liquid after superparamagnetism microballoon, reduce tunning to the feedback inhibition of producing strains, and do not add compared with superparamagnetism microballoon, capromycin output increased 24%; After complex optimum, fermentation unit improves 73%, improves production capacity, reduces the discharge of energy consumption and waste.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.Experiment material agent used in following embodiment is industrial raw material if no special instructions; Reagent is purchased from Beijing Chemical Plant, and superparamagnetism microballoon is provided by Suzhou Nano-Micro Bio-technology Co., Ltd..
Embodiment 1,5L Fermentation fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 26 DEG C, 220rpm, cultivation 72h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 20.0 grams, yeast powder 10.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 2.0 grams, 0.5 gram, sodium-chlor, 4.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH6.5,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial strain of numbering NRRL2773.
Fermention medium forms:
W-Gum 60.0 grams, soybean cake powder 60.0 grams, 12.0 grams, ammonium sulfate, corn steep liquor 12.0 grams, potassium primary phosphate 12.0 grams, 6.0 grams, sodium-chlor, 12.0 grams, calcium carbonate, add tap water and be settled to 3000mL, pH6.5,121 DEG C of sterilizing 30min, 5L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:1.2vvm, and charge amount is 3L, inoculates after seed culture 72h, inoculum size is 10%, culture temperature 29 DEG C, rotating speed 300r/min, fermentation culture adds Methionin after starting 24 hours, and often liter of fermented liquid adds 0.5g, total fermentation time 120h.
The results are shown in Table 1, add precursor substance Methionin during the fermentation, after fermentation ends, the fermentation unit measuring capromycin is 9881ug/ml.
Embodiment 2,2L Fermentation fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 64h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 40.0 grams, yeast powder 5.0 grams, 3.0 grams, ammonium sulfate, corn steep liquor 8.0 grams, 1.0 grams, sodium-chlor, 3.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial classification of bacterium numbering NRRL2773.
Fermention medium forms:
W-Gum 30.0 grams, cottonseed flour 20.0 grams, yeast powder 5.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 8.0 grams, potassium primary phosphate 2.0 grams, 1.0 grams, sodium-chlor, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min, 2L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:0.8vvm, charge amount is 1L, inoculate after seed culture 64h, inoculum size is 8%, culture temperature 29 DEG C, rotating speed is 400r/min, starts to add Methionin in 56 hours in fermentation culture, often liter of fermented liquid adds 1.5g, total fermentation time 144h.
After fermentation ends, the fermentation unit of capromycin is 11355ug/ml after testing.Concrete outcome is in table 1.
Embodiment 3,2L automatic jug fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 30 DEG C, 220rpm, cultivation 48h obtain seed liquor;
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 4.0 grams, 2.0 grams, sodium-chlor, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH6.5,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial classification of bacterium numbering NRRL2773.
Fermention medium forms:
W-Gum 30.0 grams, cottonseed flour 20.0 grams, yeast powder 5.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 8.0 grams, potassium primary phosphate 2.0 grams, 1.0 grams, sodium-chlor, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min, 2L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:1.1vvm, charge amount is 1L, inoculate after seed culture 48h, inoculum size is 7%, and culture temperature 29 DEG C, 400rpm stir, and fermentation culture adds Methionin after starting 36 hours, often liter of fermented liquid adds 2g, fermentation culture starts within 48 hours, to add the sodium form superparamagnetism microballoon that particle diameter is 5um, and often liter of fermented liquid add-on is 20g, total fermentation time 144h.
After fermentation ends, the superparamagnetism microballoon in magnetic resolution fermented liquid, adds the capromycin of the 1mol/L aqueous sulfuric acid wash-out absorption of 120ml, measures the capromycin in elutriant and fermented liquid respectively after recovery.The fermentation unit of detection computations capromycin is 14149ug/ml.Concrete outcome is in table 1.
Embodiment 4,5L tank fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 64h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 20.0 grams, yeast powder 10.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 2.0 grams, 1.0 grams, sodium-chlor, 4.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial classification of bacterium numbering NRRL2773.
Fermention medium forms:
W-Gum 60.0 grams, cottonseed flour 60.0 grams, yeast powder 15.0 grams, 9.0 grams, ammonium sulfate, corn steep liquor 18.0 grams, potassium primary phosphate 9.0 grams, 6.0 grams, sodium-chlor, 15.0 grams, calcium carbonate, add tap water and be settled to 3000mL, pH6.5,121 DEG C of sterilizing 30min, 5L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:1.1vvm, loading amount is 3L, inoculate after seed culture 64h, inoculum size is 8%, and culture temperature 27 DEG C, rotating speed are 300r/min, add fermentating liquid volume Methionin after fermentation culture starts 56 hours, often liter of fermented liquid adds 1g, fermentation culture starts within 72 hours, to add the sodium form superparamagnetism microballoon that particle diameter is 30um, and often liter of fermented liquid add-on is 10g, total fermentation time 144h.
After fermentation ends, magnetic resolution reclaims the superparamagnetism microballoon in fermented liquid, adds the capromycin of the 1mol/L aqueous hydrochloric acid wash-out absorption of 60ml, measure the capromycin in elutriant and fermented liquid respectively after recovery.The fermentation unit calculating capromycin is 14125ug/ml.Concrete outcome is in table 1.
Comparative example 1,5L Fermentation fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 26 DEG C, 220rpm, cultivation 72h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 20.0 grams, yeast powder 10.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 2.0 grams, 0.5 gram, sodium-chlor, 4.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH6.5,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial classification of bacterium numbering NRRL2773.
Fermention medium forms:
W-Gum 60.0 grams, soybean cake powder 60.0 grams, 12.0 grams, ammonium sulfate, corn steep liquor 12.0 grams, potassium primary phosphate 12.0 grams, 6.0 grams, sodium-chlor, 12.0 grams, calcium carbonate, add tap water and be settled to 3000mL, pH6.5,121 DEG C of sterilizing 30min fermentation culture, 5L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:1.2vvm, and loading amount is 3L, inoculates after seed culture 72h, inoculum size is 10%, culture temperature 29 DEG C, rotating speed 300r/min, total fermentation time 120h.
After fermentation ends, the fermentation unit of capromycin is 8166ug/ml, the results are shown in Table 1.
Comparative example 2,2L Fermentation fermentation are for capromycin
The seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 64h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 40.0 grams, yeast powder 5.0 grams, 3.0 grams, ammonium sulfate, corn steep liquor 8.0 grams, 1.0 grams, sodium-chlor, 3.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
Capreomycin-producing Streptomyces Capreolus of the present invention is the bacterial classification of bacterium numbering NRRL2773.
Fermention medium forms:
W-Gum 30.0 grams, cottonseed flour 20.0 grams, yeast powder 5.0 grams, 2.0 grams, ammonium sulfate, corn steep liquor 8.0 grams, potassium primary phosphate 2.0 grams, 1.0 grams, sodium-chlor, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min, 2L Fermentation fermentation culture.
Fermentation condition: tank pressure 0.05 ± 0.01Mpa, ventilation ratio are 1:0.8vvm, and loading amount is 1L, inoculates after seed culture 64h, inoculum size is 8%, culture temperature 29 DEG C, rotating speed 400r/min, total fermentation time 144h.
After fermentation ends, the fermentation unit of capromycin is 9232ug/ml after testing, the results are shown in Table 1.
Table 1 nitrogenous source, precursor substance and superparamagnetism microballoon prepare capromycin contrast to microorganism
Results contrast in table 1 is calculated known:
Organic nitrogen source adopts cottonseed flour and yeast powder, compares employing soybean cake powder, and fermentation unit improves (9232-8166)/8166=13%, or (11355-9881)/9881=15%.
Add precursor substance, fermentation unit improves (9881-8166//8166=21%, or (11355-9232)/9232=23%.
Add superparamagnetism microballoon in fermentation, fermentation unit improves (14149-11355)/11355=25%.Or (14125-11355)/11355=24%.
After complex optimum, i.e. embodiment 3(or 4) compared with comparative example 1, fermentation unit improves (14149-8166)/8166=73%.

Claims (6)

1. a method for fermentative production capromycin, is characterized in that: comprise the following steps:
A, prepare the seed liquor of curling streptomycete bacterial strain
The slant culture of curling streptomycete bacterial strain or spore liquid are inoculated in seed culture medium, 26-30 DEG C, cultivation 48-72h acquisition seed liquor;
Wherein said seed culture medium obtains by the following method: sucrose 20.0-40.0 gram, yeast powder 5.0-10.0 gram, ammonium sulfate 2.0-3.0 gram, corn steep liquor 2.0-8.0 gram, sodium-chlor 0.5-2.0 gram, calcium carbonate 3.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min;
B, prepare the fermented liquid of curling streptomycete bacterial strain
Above-mentioned seed liquor is inoculated in fermention medium with the inoculum size of volume percent 7-10%, in shaking flask or fermentation cylinder for fermentation, ferments between 24-56 hour and add precursor substance Methionin; Total fermentation time 120-144 hour, culture temperature 27-29 DEG C, obtain fermented liquid;
Wherein said fermention medium obtains by the following method: W-Gum 20.0-30.0 gram, cottonseed flour 0-20.0 gram, soybean cake powder 0-20.0 gram, yeast powder 0-5.0 gram, ammonium sulfate 2.0-4.0 gram, corn steep liquor 4.0-8.0 gram, potassium primary phosphate 2.0-4.0 gram, sodium-chlor 1.0-2.0 gram, calcium carbonate 4.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
2. method according to claim 1, the tank pressure that ferments in described step b is 0.05 ± 0.01Mpa, ventilation ratio is 1:0.8-1.2vvm, stirring velocity is 300-400rpm.
3. method according to claim 1, in described step b, precursor substance addition adds 0.5-2g for often liter of fermented liquid.
4. the method according to any one of claim 1-3, ferments in step b between 48-72 hour and adds the superparamagnetism microballoon after sterilizing.
5. method according to claim 4, described superparamagnetism microspherulite diameter is 5-30um, and size distribution is single dispersing, and the functional group on surface is the sodium form structure after sodium hydroxide solution process.
6. method according to claim 5, the addition of superparamagnetism microballoon adds 10-20g for often liter of fermented liquid.
CN201610097987.2A 2016-02-23 2016-02-23 Method for producing capreomycin by fermentation Pending CN105506042A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201610097987.2A CN105506042A (en) 2016-02-23 2016-02-23 Method for producing capreomycin by fermentation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201610097987.2A CN105506042A (en) 2016-02-23 2016-02-23 Method for producing capreomycin by fermentation

Publications (1)

Publication Number Publication Date
CN105506042A true CN105506042A (en) 2016-04-20

Family

ID=55714332

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201610097987.2A Pending CN105506042A (en) 2016-02-23 2016-02-23 Method for producing capreomycin by fermentation

Country Status (1)

Country Link
CN (1) CN105506042A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113755548A (en) * 2021-09-02 2021-12-07 河北圣雪大成制药有限责任公司 Method for improving fermentation level of polymyxin B

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3143468A (en) * 1962-05-25 1964-08-04 Lilly Co Eli Capreomycin and its preparation
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
CN103105446A (en) * 2011-11-15 2013-05-15 南昌大学 Method for enriching beta-lactam penicillins antibiotics in liquid milk
CN105002244A (en) * 2015-07-30 2015-10-28 宁夏泰瑞制药股份有限公司 Culture medium and culture method for producing capreomycin through Streptomyces capreolus fermentation
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

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3143468A (en) * 1962-05-25 1964-08-04 Lilly Co Eli Capreomycin and its preparation
CN103076415A (en) * 2011-11-15 2013-05-01 南昌大学 Method for enriching nature tetracycline antibiotics in aquatic products
CN103105446A (en) * 2011-11-15 2013-05-15 南昌大学 Method for enriching beta-lactam penicillins antibiotics in liquid milk
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
CN105002244A (en) * 2015-07-30 2015-10-28 宁夏泰瑞制药股份有限公司 Culture medium and culture method for producing capreomycin through Streptomyces capreolus fermentation
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 (2)

* Cited by examiner, † Cited by third party
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》 *
梁淑芳 等: "卷曲霉素产生菌的改良", 《应用与环境生物学报》 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN113755548A (en) * 2021-09-02 2021-12-07 河北圣雪大成制药有限责任公司 Method for improving fermentation level of polymyxin B

Similar Documents

Publication Publication Date Title
CN109439701B (en) Method for preparing ergothioneine by biosynthesis and fermentation medium
Ryu et al. Optimization of erythritol production by Candida magnoliae in fed-batch culture
Chen et al. Effects of culture redox potential on succinic acid production by Corynebacterium crenatum under anaerobic conditions
US20240102058A1 (en) Caproate-producing bacterium with multiple substrate utilization capabilities and its applications
WO2020037998A1 (en) Recombinant pseudomonas plecoglossicida for producing l-xylose and application thereof
Yang et al. Production of erythritol from glucose by an osmophilic mutant of Candida magnoliae
CN105483064A (en) Method for fermentation production of bacitracin
CN100363490C (en) Metabolically engineered micro-organisms having reduced production of undesired metabolic products
Lan et al. Isolation and characterization of a newly identified Clostridium butyricum strain SCUT343-4 for 1, 3-propanediol production
CN103276019B (en) Method for promoting lycopene synthesis in blakeslea trispora
Qu et al. Enhanced saccharification of cellulose and sugarcane bagasse by Clostridium thermocellum cultures with Triton X-100 and β-glucosidase/Cellic® CTec2 supplementation
CN101845407B (en) Actinobacillus and method for producing succinic acid
Guo et al. Energy-and cost-effective non-sterilized fermentation of 2, 3-butanediol by an engineered Klebsiella pneumoniae OU7 with an anti-microbial contamination system
CN105524961A (en) Method for producing amphotericin B through fermentation
NZ566781A (en) Method of culturing thermophilic microorganisms suitable for the production of ethanol
VENKATESWARAN et al. The Clostridium thermocellum-Clostridium thermosaccharolyticum ethanol production process: nutritional studies and scale-down
CN112358986B (en) Clostridium butyricum and application thereof in production of 1,3-propylene glycol through immobilized fermentation
CN112852656B (en) Escherichia coli engineering bacterium for producing glycollic acid by using glucose and acetic acid dual-carbon source
CN104277989B (en) One plant of Saccharomyces cerevisiae and its application in fermenting and producing DPN
CN105506041A (en) Method for producing norvancomycin by fermentation
CN105506042A (en) Method for producing capreomycin by fermentation
CN115637276B (en) Method for producing tetrahydropyrimidine by using halomonas strain
WO2019127829A1 (en) Fermentation production method for oxidized coenzyme q10, and high-content oxidized coenzyme q10 prepared therefrom
Cheng et al. Strain isolation and study on process parameters for xylose-to-xylitol bioconversion
CN102747114A (en) Method for regulating recombinant escherichia coli metabolism by using transient anaerobic fermentation

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