CN106755221A - A kind of preparation method of MFG parent nucleus FR179642 - Google Patents
A kind of preparation method of MFG parent nucleus FR179642 Download PDFInfo
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- 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
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Abstract
The present invention provides a kind of MFG parent nucleus producing strains, and by adding MFG precursor during the fermentation, the method for obtaining MFG parent nucleus deacylated to MFG precursor belongs to field of biological pharmacy.In the present invention, MFG parent nucleus FR179642 bacterial strains ferment in the medium, MFG precursor FR901379 and the method for passing through multistage temperature control are added at twice during the fermentation, so as to improve the yield of MFG parent nucleus FR179642；The acidified broken wall treatment of zymotic fluid, with Solvent Extract methods, macroporous resin adsorption parses to obtain desorbed solution, then obtains MFG parent nucleus powder by crystallization by desorbed solution.The present invention can be used to synthesize MFG parent nucleus FR179642, and operation sequence is simple, method stabilization, favorable reproducibility.
Description
Technical field
The present invention relates to biological pharmacy technical field, more particularly to a kind of preparation side of MFG parent nucleus FR179642
Method.
Background technology
In recent years, with the extensive application of extensive pedigree antibiotic and immunodepressant, chemotherapy of tumors, organ transplant and Chinese mugwort
Spreading for disease is grown, the crowd for causing immune system to be suppressed is on the increase, the incidence of disease of deep fungal infection starts to present
Obvious ascendant trend, and with the utilization of antifungal drug, the drug resistance of fungi is also more and more stronger so that antifungal drug
Using there is swift and violent increased trend.Therefore, the medicine of anti-deep fungal infection has turned into one of study hotspot of anti-infectious agent, day
Benefit causes the concern of people.
Echinocandin is the new antifungal of a class, and the mechanism of action with current clinical conventional antifungal drug is obvious not
Together, it is β -1, the inhibitor of 3- glucan synthases acts on the cell membrane of fungi.Cell membrane is fungi and mammalian cell
Between maximum difference, so the synthesis of interference cell wall, influence the integrality of cell membrane that fungal cell will be caused dead and right
People's fanout free region, therefore side effect is smaller, security is higher.
MFG is a kind of water-soluble semi synthesis echinocandin antifungal agent thing, and FR179642 is synthesis meter Ka Fen
Net parent nucleus, CAS accession number：168110-44-9, chemical formula：C35H52N8O20S, molecular weight：936.90.The chemistry knot of FR179642
Structure formula is as follows：
At present, the report in terms of relevant FR179642 synthesis or preparation is detected in publication number KR20140039753's (A)
In South Korea's patent of invention.It passes through intermediate FR901379 through deacetylation, and is prepared by starting material of FR901379
FR179642.Specific syntheti c route is as follows：
But, there is preparation cost defect high in above-mentioned prior art.Meanwhile, there is no in the prior art for FR179642
Relevant report prepared by fermentation, belongs to the blanking technique of current industry.
The content of the invention
It is an object of the invention to disclose a kind of preparation method of MFG parent nucleus FR179642,
For achieving the above object, the invention provides the preparation method of MFG parent nucleus FR179642, including with
Lower step：
Step (1), a small amount of MFG parent nucleus FR179642 producing strains spore suspensions are taken, are inoculated on seed culture medium,
In at 24~28 DEG C, 40~50h is cultivated；
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, reached when the bacterium in fermentation tank is dense
MFG precursor FR901379 is added after to stable state, MFG precursor is added after continuing 40~60h of fermentation
Dual-stage temperature control, fermentation completion are used in FR901379, the step (2) to be carried out putting tank extracting zymotic fluid；
Step (3), make the pH of zymotic fluid in 3~4 to adding acid in zymotic fluid, then plate-frame filtering adds polar solvent
Concentration is stripped, concentrate is obtained；
Step (4), by concentrate through macroporous resin adsorption, parsed using polar solvent, obtain MFG parent nucleus
FR179642 desorbed solutions；
Step (5), MFG parent nucleus FR179642 desorbed solutions is crystallized, drying, obtain in powdered MFG
Parent nucleus FR179642.
As a further improvement on the present invention, the component of the seed culture medium in the step (1) is：Glucose 20~
35g/L, 5~15g/L of dusty yeast, 5~15g/L of soybean cake powder, 0.5~1.5g/L of dipotassium hydrogen phosphate, 0.1~1.0g/ of sodium chloride
L, 0.1~1.0g/L of magnesium sulfate, 0.1~0.5g/L of calcium carbonate, balance of water.
As a further improvement on the present invention, the component of the fermentation medium in the step (2) is：Glucose 5~
15g/L, 70~90g/L of dusty yeast, 6~10g/L of soybean cake powder, 1~5g/L of dipotassium hydrogen phosphate, 1~5g/L of sodium chloride, magnesium sulfate
0.2~0.4g/L, 0.1~0.5g/L of calcium carbonate, balance of water.
As a further improvement on the present invention, in the step (2), MFG precursor FR901379's in fermentation tank
Concentration stabilization is between 500~2000 μ g/mL.
As a further improvement on the present invention, in the step (2), in the step (2), from original state to fermentation tank
In the dense fermentation stage for reaching stable state of interior bacterium, the temperature control in fermentation tank is at 30~34 DEG C；Bacterium in fermentation tank is dense
Stable state is reached into putting the tank stage, and the temperature control in fermentation tank is at 26~30 DEG C.
As a further improvement on the present invention, the acid in the step (3) is selected from the dilute sulfuric acid that concentration is 5~10wt%, 5
The dilute sulfuric acid of~10wt% or the phosphoric acid,diluted of 5~10wt%.
As a further improvement on the present invention, the polar solvent in the step (4) is selected from the alcohol of C1~C4；The step
(4) macroreticular resin in is selected from HP20 macroreticular resins, D1300 macroreticular resins or SP207 macroreticular resins.
As a further improvement on the present invention, the crystallization mode in the step (5) is that solvent settles crystallization or concentrates
Crystallization.
As a further improvement on the present invention, the used solvent of the solvent sedimentation crystallization is selected from petroleum ether, n-hexane
Or hexamethylene.
As a further improvement on the present invention, the drying mode in the step (5) is vacuum drying, vacuum drying dry
Dry temperature is 40~50 DEG C, and vacuum is 0.1~0.5MPa.
Compared with prior art, the beneficial effects of the invention are as follows：In the present invention, MFG parent nucleus FR179642 bacterial strains
Ferment in the medium, add MFG precursor FR901379 and the side for passing through multistage temperature control at twice during the fermentation
Method, improves the yield of MFG parent nucleus FR179642；The acidified broken wall treatment of zymotic fluid, with Solvent Extract methods, macropore
Resin adsorption parses to obtain desorbed solution, then obtains MFG parent nucleus powder by crystallization by desorbed solution.The present invention can be used for
Synthesis MFG parent nucleus FR179642, and operation sequence is simple, method stabilization, favorable reproducibility.
Specific embodiment
With reference to each implementation method, the present invention is described in detail, but it should explanation, these implementation methods are simultaneously
Non- limitation of the present invention, those of ordinary skill in the art are according in these implementation method institutes works energy, method or structure
Equivalent transformation or replacement, belong within protection scope of the present invention.
Unless there is specified otherwise in specification, component, the raw material in each embodiment in the present invention are pure using analysis
Rank.In addition, " g " in each embodiment is unit of weight " gram "；" h " is chronomere's " hour "；" ml " is volume unit " milli
Rise "；" room temperature " is 23 DEG C；“BV/H”：The expression of space flow speed, that is, refer to the unit interval (hour) to flow through unit volume tree in post
The average liquid measure of fat.“HPLC”：High performance liquid chromatography.
Embodiment one：
The preparation method of MFG parent nucleus FR179642, comprises the following steps.
Step (1), a small amount of MFG parent nucleus FR179642 producing strains spore suspensions are taken, are inoculated on seed culture medium,
In at 24 DEG C, 50h is cultivated.Component in the seed culture medium is (unit：g/L)：Glucose 20, dusty yeast 5, soybean cake powder 5,
Dipotassium hydrogen phosphate 0.5, sodium chloride 0.1, magnesium sulfate 0.1, calcium carbonate 0.1, balance of water.
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, in culture 60 at 24~28 DEG C~
70h.MFG precursor FR901379 is added after the bacterium in fermentation tank is dense reaches stable state, after continuing fermentation 40h, is added
MFG precursor FR901379.In the two fermentation stages, from original state to fermentation tank in bacterium it is dense reach stabilization shape
First fermentation stage of state, the temperature control in fermentation tank is at 30 DEG C.Bacterium from fermentation tank is dense to reach stable state to putting
In second fermentation stage of tank, the temperature control in fermentation tank is at 26 DEG C.Component in the fermentation medium is (unit：g/
L)：Glucose 5, dusty yeast 70, soybean cake powder 6, dipotassium hydrogen phosphate 1, sodium chloride 1, magnesium sulfate 0.2, calcium carbonate 0.1 is balance of
Water.In the present embodiment, in second fermentation stage, the concentration stabilization of the MFG precursor FR901379 in fermentation tank exists
500 μ g/mL or so.
Step (3), fermentation completion extracted after putting tank.When extracting zymotic fluid, it is to addition concentration in fermentation tank first
The dilute sulfuric acid of 5wt%, so that the system of zymotic fluid is acidified to pH in 3 in fermentation tank, after crossing sheet frame, plus methyl alcohol is used as polar solvent
Extracting concentration, obtains MFG parent nucleus FR179642 concentrates.
Step (4), MFG parent nucleus FR179642 concentrates are entered using the macroporous resin adsorption post for being filled with HP20
Row absorption.Adsorption flow rate controls 0.5~1.5BV/H, is washed after the completion of absorption, is 60wt% methyl alcohol as polar solvent with concentration
Parsed, parsed 1~2BV/H of flow velocity, merged active principle, obtained MFG parent nucleus FR179642 desorbed solutions.
Step (5), the crystallization of MFG parent nucleus FR179642 desorbed solutions, dry, and obtain in the MFG parent nucleus of powdery
FR179642.In the present embodiment, crystallization mode is condensing crystallizing, and crystallization temperature is 40 DEG C.Drying mode is vacuum drying, is dried
40 DEG C of temperature, vacuum drying vacuum is 0.1MPa.
MFG parent nucleus FR179642 obtained by the present embodiment normalizes purity 85.5% through HPLC detections.
Embodiment two：
The preparation method of MFG parent nucleus FR179642, comprises the following steps.
Step (1), a small amount of MFG parent nucleus FR179642 producing strains spore suspensions are taken, are inoculated on seed culture medium,
In at 28 DEG C, 40h is cultivated.Component in the seed culture medium is (unit：g/L)：Glucose 35, dusty yeast 15, soybean cake powder
15, dipotassium hydrogen phosphate 1.5, sodium chloride 1.0, magnesium sulfate 1.0, calcium carbonate 0.5, balance of water.
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, in culture 60 at 24~28 DEG C~
70h。
MFG precursor FR901379 is added after the bacterium in fermentation tank is dense reaches stable state, after continuing fermentation 60h,
Add MFG precursor FR901379.In the two fermentation stages, from original state to fermentation tank in bacterium it is dense reach it is steady
Determine first fermentation stage of state, the temperature control in fermentation tank is at 31 DEG C.Stable state is reached from the bacterium of fermentation tank is dense extremely
Put in second fermentation stage of tank, the temperature control in fermentation tank is at 27 DEG C.Component in the fermentation medium component is (single
Position：g/L)：Glucose 15, dusty yeast 90, soybean cake powder 10, dipotassium hydrogen phosphate 5, sodium chloride 5, magnesium sulfate 0.4, calcium carbonate 0.5,
Balance of water.In the present embodiment, in second fermentation stage, the MFG precursor concentration stabilization in fermentation tank is 2000
μ g/mL or so.
Step (3), fermentation completion extracted after putting tank.When extracting zymotic fluid, it is to addition concentration in fermentation tank first
The watery hydrochloric acid of 5wt%, so that the system of zymotic fluid is acidified to pH in 3 in fermentation tank, after crossing sheet frame, plus ethanol is used as polar solvent
Concentration is stripped, MFG parent nucleus FR179642 concentrates are obtained.
Step (4), concentrate is adsorbed using the macroporous resin adsorption post for being filled with SP207.Adsorption flow rate is controlled
0.5~1.5BV/H, washes after the completion of absorption, is parsed as polar solvent with the ethanol that concentration is 60wt%, parses flow velocity
1~2BV/H, merges active principle, obtains MFG parent nucleus FR179642 desorbed solutions.
Step (5), by the crystallization of MFG parent nucleus FR179642 desorbed solutions, dry, obtain female in the MFG of powdery
Core FR179642.Crystallization mode is 50 DEG C of condensing crystallizings.Drying mode is vacuum drying, 50 DEG C of drying temperature, vacuum is
0.5MPa。
MFG parent nucleus FR179642 obtained by the present embodiment normalizes purity 85.7% through HPLC detections.
Embodiment three：
The preparation method of MFG parent nucleus FR179642, comprises the following steps.
Step (1), a small amount of MFG parent nucleus FR179642 producing strains spore suspensions are taken, be inoculated on culture medium, in
At 26 DEG C, 48h is cultivated.Component in the seed culture medium is (unit：g/L)：Glucose 25, dusty yeast 10, soybean cake powder 10,
Dipotassium hydrogen phosphate 1, sodium chloride 0.5, magnesium sulfate 0.5, calcium carbonate 0.2, balance of water.
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, in culture 60 at 24~28 DEG C~
70h。
MFG precursor FR901379 is added after the bacterium in fermentation tank is dense reaches stable state.After continuing fermentation 50h,
Add MFG precursor FR901379.In the two fermentation stages, from original state to fermentation tank in bacterium it is dense reach it is steady
Determine first fermentation stage of state, the temperature control in fermentation tank is at 33 DEG C.Bacterium from fermentation tank is dense to reach stable state
Into putting second fermentation stage of tank, the temperature control in fermentation tank is at 28 DEG C.Component in the fermentation medium is (single
Position：g/L)：Glucose 10, dusty yeast 80, soybean cake powder 8, dipotassium hydrogen phosphate 2, sodium chloride 2, magnesium sulfate 0.3, calcium carbonate 0.3,
Balance of water.In the present embodiment, in second fermentation stage, the stabilization of the MFG precursor FR901379 in fermentation tank
In 700 μ g/mL.
Carry out putting tank extraction after the completion of step (3), fermentation.When extracting zymotic fluid, it is to addition concentration in fermentation tank first
The watery hydrochloric acid of 5wt%, so that the system of zymotic fluid is acidified to pH in 3.5 in fermentation tank, plate-frame filtering, then to adding in fermentation tank
Plus the methyl alcohol that concentration is 60wt% is stripped concentration as polar solvent, obtains MFG parent nucleus FR179642 desorbed solutions.
Step (4), MFG parent nucleus FR179642 desorbed solutions are carried out using D1300 macroporous resin adsorptions post is filled with
Absorption, is parsed with the methyl alcohol that concentration is 60wt% as polar solvent, parses 1~2BV/H of flow velocity, merges active principle,
It is concentrated to give MFG parent nucleus FR179642 desorbed solutions.
Step (5), by the crystallization of MFG parent nucleus FR179642 desorbed solutions, filtering is dried, obtained in powdered rice card
Fragrant net parent nucleus FR179642.Crystallization mode is that solvent settles crystallization, and is specially：To MFG parent nucleus FR179642 desorbed solutions
15~20 times of hexamethylenes of volume of middle addition.In the present embodiment, drying mode is vacuum drying, 40 DEG C of drying temperature, vacuum
It is 0.2MPa to spend.
MFG parent nucleus FR179642 obtained by the present embodiment normalizes purity 86.1% through HPLC detections.
Example IV：
The preparation method of MFG parent nucleus FR179642, comprises the following steps.
Step (1), a small amount of FR179642 producing strains spore suspensions are taken, be inoculated on culture medium, at 26 DEG C, culture
44h.Component in the seed culture medium is (unit：g/L)：Glucose 20, dusty yeast 15, soybean cake powder 5, dipotassium hydrogen phosphate
0.5, sodium chloride 0.1, magnesium sulfate 0.1, calcium carbonate 0.1, balance of water.
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, in cultivating 60 at 24~28 DEG C
~70h.
MFG precursor FR901379 is added after the bacterium in fermentation tank is dense reaches stable state, after continuing fermentation 55h,
Add MFG precursor FR901379.In the two fermentation stages, from original state to fermentation tank in bacterium it is dense reach it is steady
Determine a fermentation stage of state, the temperature control in fermentation tank is at 32 DEG C.Bacterium from fermentation tank is dense to reach stable state extremely
Put in second fermentation stage of tank, the stability contorting in fermentation tank is at 29 DEG C.Component in the fermentation medium component is (single
Position：g/L)：Glucose 5, dusty yeast 70, soybean cake powder 6, dipotassium hydrogen phosphate 1, sodium chloride 1, magnesium sulfate 0.2, calcium carbonate 0.1 is remaining
It is water to measure.In this example, in second fermentation stage, the concentration stabilization of the MFG precursor FR901379 in fermentation tank
In 800 μ g/mL or so.
Step (3), fermentation completion extracted after putting tank.When extracting zymotic fluid, it is to addition concentration in fermentation tank first
The phosphoric acid,diluted of 5wt%, so that the system of zymotic fluid is acidified to pH in 3 in fermentation tank, after crossing sheet frame, plus ethanol is used as polar solvent
Extracting concentration, obtains MFG parent nucleus FR179642 concentrates.
Step (4), MFG parent nucleus FR179642 concentrates are entered using the macroporous resin adsorption post for being filled with D1300
Row absorption.Adsorption flow rate controls 0.5~1.5BV/H, is washed after the completion of absorption, molten as polarity with the ethanol that concentration is 60wt%
Agent is parsed, and parses 1~2BV/H of flow velocity, merges active principle, obtains MFG parent nucleus FR179642 desorbed solutions.
Step (5), by the crystallization of MFG parent nucleus FR179642 desorbed solutions, dry, obtain in powdered MFG
Parent nucleus FR179642.Crystallization mode is that solvent settles crystallization, and is specially：Add in MFG parent nucleus FR179642 desorbed solutions
Enter 15~20 times of n-hexanes of volume.Crystallization and filtration is simultaneously dried.Specifically, drying mode is vacuum drying, drying temperature 40
DEG C, vacuum is 0.3MPa.
MFG parent nucleus FR179642 obtained by the present embodiment normalizes purity 85.7% through HPLC detections.
Embodiment five：
The preparation method of MFG parent nucleus FR179642, comprises the following steps.
Step (1), a small amount of FR179642 producing strains spore suspensions are taken, be inoculated on culture medium, at 26 DEG C, culture
40h.Component in the seed culture medium is (unit：g/L)：Glucose 35, dusty yeast 15, soybean cake powder 15, dipotassium hydrogen phosphate
1.5, sodium chloride 1.0, magnesium sulfate 1.0, calcium carbonate 0.5, balance of water.
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, in cultivating 60 at 24~28 DEG C
~70h.
MFG precursor FR901379 is added after the bacterium in fermentation tank is dense reaches stable state, after continuing fermentation 60h,
Add MFG precursor FR901379.In the two fermentation stages, from original state to fermentation tank in bacterium it is dense reach it is steady
Determine a fermentation stage of state, the temperature control in fermentation tank is at 34 DEG C.Bacterium from fermentation tank is dense to reach stable state extremely
Put in second fermentation stage of tank, the stability contorting in fermentation tank is at 30 DEG C.Component in the fermentation medium component is (single
Position：g/L)：Glucose 15, dusty yeast 90, soybean cake powder 10, dipotassium hydrogen phosphate 5, sodium chloride 5, magnesium sulfate 0.4, calcium carbonate 0.5,
Balance of water.In this example, in second fermentation stage, the concentration of the MFG precursor FR901379 in fermentation tank is steady
It is scheduled on 2000 μ g/mL or so.
Step (3), fermentation completion extracted after putting tank；When extracting zymotic fluid, it is to addition concentration in fermentation tank first
The watery hydrochloric acid of 5wt%, so that the system of zymotic fluid is acidified to pH in 3 in fermentation tank, after crossing sheet frame, plus methyl alcohol is used as polar solvent
Extracting concentration, obtains MFG parent nucleus FR179642 concentrates.
Step (4), the MFG parent nucleus FR179642 concentrates for obtaining use the macroporous resin adsorption for being filled with D1300
Post is adsorbed.Adsorption flow rate controls 0.5~1.5BV/H, is washed after the completion of absorption, is solved with the methyl alcohol that concentration is 60wt%
Analysis, parses 1~2BV/H of flow velocity, merges active principle, obtains MFG parent nucleus FR179642 desorbed solutions.
Step (5), by the crystallization of MFG parent nucleus FR179642 desorbed solutions, dry, obtain in powdered MFG
Parent nucleus FR179642.Crystallization mode is that solvent settles crystallization, and is specially：Add in MFG parent nucleus FR179642 desorbed solutions
Enter 15~20 times of petroleum ethers of volume.Crystallization and filtration is simultaneously dried.Specifically, drying mode is vacuum drying, drying temperature 40
DEG C, vacuum is 0.4MPa.
MFG parent nucleus FR179642 obtained by the present embodiment normalizes purity 86.3% through HPLC detections.
Those listed above is a series of to be described in detail only for feasibility implementation method of the invention specifically
Bright, they simultaneously are not used to limit the scope of the invention, all equivalent implementations made without departing from skill spirit of the present invention
Or change should be included within the scope of the present invention.
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, Er Qie
In the case of without departing substantially from spirit or essential attributes of the invention, the present invention can be in other specific forms realized.Therefore, no matter
From the point of view of which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the present invention is by appended power
Profit requires to be limited rather than described above, it is intended that all in the implication and scope of the equivalency of claim by falling
Change is included in the present invention.
Moreover, it will be appreciated that although the present specification is described in terms of embodiments, not each implementation method is only wrapped
Containing an independent technical scheme, this narrating mode of specification is only that for clarity, those skilled in the art should
Specification an as entirety, the technical scheme in each embodiment can also be formed into those skilled in the art through appropriately combined
May be appreciated other embodiment.
Claims (10)
1. the preparation method of MFG parent nucleus FR179642, it is characterised in that comprise the following steps：
Step (1), a small amount of MFG parent nucleus FR179642 producing strains spore suspensions are taken, be inoculated on seed culture medium, in 24
At~28 DEG C, 40~50h is cultivated；
Step (2), seed culture medium is inoculated in fermentation tank and fermentation medium is added, when the bacterium in fermentation tank is dense reach it is steady
MFG precursor FR901379 is added after determining state, MFG precursor FR901379, institute are added after continuing 40~60h of fermentation
State step (2) middle using dual-stage temperature control, fermentation completion carries out putting tank extracting zymotic fluid；
Step (3), make the pH of zymotic fluid in 3~4 to adding acid in zymotic fluid, plate-frame filtering, then adding polar solvent is carried out
Extracting concentration, obtains concentrate；
Step (4), by concentrate through macroporous resin adsorption, parsed using polar solvent, obtain MFG parent nucleus FR179642
Desorbed solution；
Step (5), MFG parent nucleus FR179642 desorbed solutions is crystallized, drying, obtain in powdered MFG parent nucleus
FR179642。
2. preparation method according to claim 1, it is characterised in that the component of the seed culture medium in the step (1)
For：20~35g/L of glucose, 5~15g/L of dusty yeast, 5~15g/L of soybean cake powder, 0.5~1.5g/L of dipotassium hydrogen phosphate, chlorination
0.1~1.0g/L of sodium, 0.1~1.0g/L of magnesium sulfate, 0.1~0.5g/L of calcium carbonate, balance of water.
3. preparation method according to claim 1, it is characterised in that the component of the fermentation medium in the step (2)
For：5~15g/L of glucose, 70~90g/L of dusty yeast, 6~10g/L of soybean cake powder, 1~5g/L of dipotassium hydrogen phosphate, sodium chloride 1
~5g/L, 0.2~0.4g/L of magnesium sulfate, 0.1~0.5g/L of calcium carbonate, balance of water.
4. preparation method according to claim 1, it is characterised in that in the step (2), the MFG in fermentation tank
The concentration stabilization of precursor FR901379 is between 500~2000 μ g/mL.
5. preparation method according to claim 1, it is characterised in that in the step (2), from original state to fermentation tank
In the dense fermentation stage for reaching stable state of interior bacterium, the temperature control in fermentation tank is at 30~34 DEG C；Bacterium in fermentation tank is dense
Stable state is reached into putting the tank stage, and the temperature control in fermentation tank is at 26~30 DEG C.
6. preparation method according to claim 1, it is characterised in that the acid in the step (3) be selected from concentration be 5~
The phosphoric acid,diluted of the dilute sulfuric acid of 10wt%, the dilute sulfuric acid of 5~10wt% or 5~10wt%.
7. preparation method according to claim 1, it is characterised in that the polar solvent in the step (4) be selected from C1~
The alcohol of C4；Macroreticular resin in the step (4) is selected from HP20 macroreticular resins, D1300 macroreticular resins or SP207 macropore trees
Fat.
8. preparation method according to claim 1, it is characterised in that crystallization mode in the step (5) is heavy for solvent
Drop crystallization or condensing crystallizing.
9. preparation method according to claim 8, it is characterised in that the used solvent of the solvent sedimentation crystallization is selected from
Petroleum ether, n-hexane or hexamethylene.
10. preparation method according to claim 1, it is characterised in that drying mode in the step (5) is dry for vacuum
Dry, vacuum drying drying temperature is 40~50 DEG C, and vacuum is 0.1~0.5MPa.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111434675A (en) * | 2019-01-11 | 2020-07-21 | 苏州纳微科技股份有限公司 | Separation and purification method of micafungin mother ring |
| CN114874919A (en) * | 2022-05-09 | 2022-08-09 | 中国科学院青岛生物能源与过程研究所 | Micafungin precursor FR901379 high-yield strain and application thereof |
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