CN102827123B - Separation and extraction process for Mevastatin - Google Patents
Separation and extraction process for Mevastatin Download PDFInfo
- Publication number
- CN102827123B CN102827123B CN201210281403.9A CN201210281403A CN102827123B CN 102827123 B CN102827123 B CN 102827123B CN 201210281403 A CN201210281403 A CN 201210281403A CN 102827123 B CN102827123 B CN 102827123B
- Authority
- CN
- China
- Prior art keywords
- bacterium slag
- toluene
- extraction
- filtrate
- cyclisation
- 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.)
- Active
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a process for purification and extraction of Mevastatin from fermentation broth. The process is characterized in that the fermentation broth containing Mevastatin is pretreated with acid and bacterial slag is subjected to blow drying. The process provided by the invention enables a production period to be shortened, cost to be reduced and yield of a crude product of Mevastatin to be increased.
Description
Technical field
The present invention relates to a kind of method of isolated or purified mevastatin or its pharmacy acceptable salt from microbial fermentation solution.
Background technology
Mevastatin (mevastatin, be called again as shown in Equation 1) compactin (compactin), ML-236B, it is a kind of 3-hydroxyl 3-methyl glutaryl coenzyme A reductase (HMG-CoA) inhibitor, obtained by the common drugmaker biochemist Endo of Japan three accidental extraction in fungi (Penicillium Citrinum) nutrient solution, its chemistry 2-Methyl Butyric Acid-{ 1S-[1 α (R) by name, 7 β, 8 β (2S, 4S), 8a β] }-1, 2, 3, 7, 8, 8a-six hydrogen-7-methyl-8{2-[4-hydroxyl-6-oxo-2H-THP trtrahydropyranyl]-ethyl }-1-naphthalene ester, concrete structure formula is as follows:
Mevastatin is first-generation statin substance, is used as now the precursor of Statins active medicine Pravastatin more, and therefore the yield of mevastatin and quality directly affect cost and the purity of Pravastatin.
In traditional separation-extraction technology of mevastatin, mostly adopting the technique of macroporous resin adsorption column chromatography, there is the problems such as complex operation, extraction cost is high, the production cycle is long, yield is low in this technique, gradually replace by the method for solvent extraction.
US6387258 (B1) adopts the method for adjust ph, reaches the object of separation and Extraction mevastatin.The separation-extraction technology of mevastatin is disclosed: pretreated fermentation liquid under (1) alkaline environment, to remove impurity in this description of the invention; (2) statin substance is extracted with hydroxy acid or lactone form under sour environment with hydrophobicity organic extractant; (3) by extraction liquid and separation of fermentative broth; (4) the concentrated organic extractant comprising statin substance, Statins forms lactone simultaneously; (5) alkali (pH7.5-10NH
4oH, 1-5% (w/w) Na
2cO
3or NaHCO
3) solution pre-treatment, crystallization.In this invention, do not comprise bacteriological filtration slag in fermentation liquor pretreatment, directly extracted in fermented liquid, consume a large amount of organic solvents; And the acid amount that acid adjustment consumes is large, with the neutralization of a large amount of alkali during aqueous phase discharge, bring pressure to environmental protection, material cost improves.Extraction yield under alkaline condition is low, brings direct impact to whole Hydrolysis kinetics yield.In specification sheets disclosed in the method, optimum mevastatin crude yield is 78%.
Li Min discloses the extraction process of dry method and wet method lixiviate two kinds of mevastatins in " mevastatin Study on extraction ".The step that dry method extracts mevastatin is filter after fermented liquid dilute acid pretreatment, wet mycelia passes through air stream drying, drying mode has air intake 140 DEG C of air stream dryings, air intake 80 DEG C of air stream dryings and air intake 160 DEG C of flash distillations three kinds, the cyclisation of mevastatin completes while air stream drying, and dried bacterium slag (water content is lower than 5%) is for the follow-up technique such as extracting.The mevastatin crude yield that dry method extraction mevastatin obtains is only 60%.And wet method lixiviate to extract the step of mevastatin be after mycelia in fermented liquid obtains after filtration, with butylacetate, lixiviate 4 hours, lixiviate ratio is 1: 7; Second time extraction time is 2 hours, and material ratio is 1: 4; The mode extracting the distillation of concentrated solution vacuum decompression is carried out, and closed loop thermal suitable in production process is 70-80 DEG C, and the time is 6 hours.Be determined by experiment the optimum solvent of lixiviate, top condition, the mevastatin crude yield obtained is only 70%.
As can be seen from the above, present mevastatin separation-extraction technology all also exists and extract the low problem of mevastatin total recovery from fermented liquid, the yield that mevastatin is lower not only can cause great waste, and its production of raising that can be invisible rises this, be unfavorable for the reduction of the statins price taking mevastatin as precursor, thus increase the economic pressures of such drug use person.
Summary of the invention
The object of this invention is to provide that a kind of yield is high, with short production cycle, the method for separation and Extraction mevastatin or its pharmacy acceptable salt from microbial fermentation solution that adapts to scale operation.
The invention before extraction bacterium slag, introduce the step drying up bacterium slag, surprisingly find that this step can improve the yield of mevastatin crude product significantly.Bacterium slag residuary water after filtration, some water-soluble hydrophilic impurity and hold volatile impurity etc., these impurity directly can affect extraction and the cyclisation efficiency in later stage, the time of prolongation cyclisation and extraction process.Bacterium slag dries up in process, some water-soluble hydrophilic impurity and hold volatile impurity etc. along with water vapour and be removed, and in addition, the bacterium slag weight after drying up reduces, and can reduce amount with an organic solvent in follow-up extraction, shortens the time of extraction cyclisation simultaneously.
Concrete, method of the present invention comprises the following steps:
(1) pre-treatment is carried out in the fermented liquid acid containing mevastatin, filter, bacterium slag dries up
Fermented liquid pH value preferred value after acidifying is 2.0-6.0; After bacterium slag dries up, water content is 5%-76%.
(2) bacterium slag and extraction solvent drop in extractor, insulation cyclisation, filtration washing
Extraction solvent is hydrophobic organic solvent, but be selected from be not limited in butylacetate, ethyl acetate, propyl acetate, ethyl formate, hexanone, methylene dichloride, chloroform, tetracol phenixin, ethylene dichloride, toluene, phenylcarbinol one or several.
The input ratio of extraction solvent and bacterium slag is, total input amount (V/W) of extraction solvent is equivalent to the 1.0-10.0 of the total input amount of bacterium slag doubly (V/W).
(3) extract filtrate concentrating, crystallization, filtering drying obtains mevastatin
Preferably, concentrate between crystallization two steps, add the organic solvent that the mevastatin solubleness of same temperature is low in concentrated solution, insulated and stirred, crystallization.Filter, dry thick wet product.
The organic solvent that mevastatin solubleness is low, is selected from one or more in acetone, butyl ester, normal hexane, ethanol.
Crystallization is filtered the filter cake obtained and is vacuumized, and soaks for several times and drains, dry thick wet product with the organic solvent that mevastatin solubleness is low.
Preferably, method of the present invention comprises the following steps:
(1) pre-treatment is carried out in the fermented liquid acid containing mevastatin, filter, bacterium slag dries up
Fermented liquid pH value after acidifying is 3.0-4.5; After bacterium slag dries up, water content is 40%-70%.
(2) bacterium slag and extraction solvent drop in extractor, insulation cyclisation, filtration washing
Extraction solvent is hydrophobic organic solvent, is selected from one or several in chloroform, tetracol phenixin, ethylene dichloride, toluene, phenylcarbinol.
The input ratio of extraction solvent and bacterium slag is, total input amount (V/W) of extraction solvent is equivalent to the 3.0-8.0 of the total input amount of bacterium slag doubly (V/W).
(3) extract filtrate concentrating, crystallization, filtering drying obtains mevastatin
Preferably, concentrate between crystallization two steps, add the organic solvent that the mevastatin solubleness of same temperature is low in concentrated solution, insulated and stirred, crystallization.Filter, dry thick wet product.
The organic solvent that mevastatin solubleness is low, is selected from one or more in acetone, butyl ester, normal hexane, ethanol.
Crystallization is filtered the filter cake obtained and is vacuumized, and soaks for several times and drains, dry thick wet product with the organic solvent that mevastatin solubleness is low.
Preferred, method of the present invention comprises the following steps:
(1) pre-treatment is carried out in the fermented liquid acid containing mevastatin, filter, bacterium slag dries up,
Fermented liquid pH value after acidifying is 3.0-4.5; After bacterium slag dries up, water content is 50%-60%.
(2) bacterium slag and extraction solvent drop in extractor, insulation cyclisation, filtration washing
Extraction solvent is hydrophobic organic solvent, is selected from one or several in chloroform, toluene, phenylcarbinol.
The input ratio of extraction solvent and bacterium slag is, total input amount (V/W) of extraction solvent is equivalent to the 3.0-8.0 of the total input amount of bacterium slag doubly (V/W).
(3) extract filtrate concentrating, crystallization, filtering drying obtains mevastatin
Preferably, concentrate between crystallization two steps, add the organic solvent that the mevastatin solubleness of same temperature is low in concentrated solution, insulated and stirred, crystallization.Filter, dry thick wet product.
The organic solvent that mevastatin solubleness is low, is selected from one or more in acetone, butyl ester, normal hexane, ethanol.
Crystallization is filtered the filter cake obtained and is vacuumized, and soaks for several times and drains, dry thick wet product with the organic solvent that mevastatin solubleness is low.
accompanying drawing explanation
Fig. 1 is the process flow sheet of the inventive method.
embodiment
Referring to specific embodiment, the present invention is described.It will be appreciated by those skilled in the art that these embodiments are only for illustration of the present invention, its scope do not limited the present invention in any way.
embodiment 1
Experiment A
The isobutyl acetate of 5L and the mixing of 5L water, after 0.5h, continuous print adds to containing in mevastatin fermented liquid (10L).PH9.0-9.6 is regulated with dense NaOH.Mixture be heated to 60 DEG C and maintain this temperature 2 hours.Organic phase is separated with aqueous phase counter-current separation method.Add sodium lauryl sulphate when being separated and help emulsifying mixt.
The pH2.0-4.5 of the fermented liquid after purifying is regulated with sulfuric acid.Continuous print adds 5L isobutyl acetate in fermented liquid, and the isobutyl acetate comprising mevastatin is separated by continuous print.
Under vacuo isobutyl acetate is concentrated to about 150mL mutually.Concentrated solution is diluted to 800mL, uses NaHCO
3washing.After washing, concentrated solution is concentrated to 150mL again.Concentrated solution is cooled to 0-5 DEG C.Mevastatin is obtained after spending the night.
Experiment B
The isobutyl acetate of 5L and the mixing of 5L water, after 0.5h, continuous print adds to containing in mevastatin fermented liquid (10L).Mixture be heated to 60 DEG C and maintain this temperature 2 hours.Organic phase is separated with aqueous phase counter-current separation method.Add sodium lauryl sulphate when being separated and help emulsifying mixt.
The pH2.0-4.5 of the fermented liquid after purifying is regulated with sulfuric acid.Continuous print adds 5L isobutyl acetate in fermented liquid, and the isobutyl acetate comprising mevastatin is separated by continuous print.
Under vacuo isobutyl acetate is concentrated to about 150mL mutually.Concentrated solution is diluted to 800mL, uses NaHCO
3washing.After washing, concentrated solution is concentrated to 150mL again.Concentrated solution is cooled to 0-5 DEG C.Mevastatin is obtained after spending the night.
Experiment C
The isobutyl acetate of 5L and the mixing of 5L water, after 0.5h, continuous print adds in mevastatin fermented liquid (10L).PH9.0-9.6 is regulated with dense NaOH.Mixture be heated to 60 DEG C and maintain this temperature 2 hours.Organic phase is separated with aqueous phase counter-current separation method.Add sodium lauryl sulphate when being separated and help emulsifying mixt.
The pH2.0-4.5 of the fermented liquid after purifying is regulated with sulfuric acid.Continuous print adds 5L isobutyl acetate in fermented liquid, and the isobutyl acetate comprising mevastatin is separated by continuous print.
Under vacuo isobutyl acetate is concentrated mutually.Concentrated solution is cooled to 0-5 DEG C.Mevastatin crude product is obtained after spending the night.
Experiment D
With reference to " mevastatin Study on extraction "; Li Min; In University Of Tianjin's master thesis in 2007, dry method extracts the flow operations of mevastatin.
After mycelia in fermented liquid (10L) obtains after filtration, with butylacetate, at temperature 35-40 DEG C, lixiviate 4 hours, lixiviate ratio is 1: 7; Second time extraction time is 2 hours, and material ratio is 1: 4; The mode extracting the distillation of concentrated solution vacuum decompression is carried out, and closed loop thermal suitable in production process is 70-80 DEG C, and the time is 6 hours, and the butylacetate comprising mevastatin is separated by continuous print, and concentrated solution is cooled to 0-5 DEG C.Mevastatin crude product is obtained after spending the night.
Experiment E
With reference to " mevastatin Study on extraction "; Li Min; In University Of Tianjin's master thesis in 2007, dry method extracts the flow operations of mevastatin.
After mycelia in fermented liquid (10L) obtains after filtration, with the air stream drying of temperature higher than 80 DEG C, dry mycelia butylacetate, at temperature 35-40 DEG C, lixiviate 4 hours, lixiviate ratio is 1: 7; Second time extraction time is 2 hours, and material ratio is 1: 4; The mode extracting the distillation of concentrated solution vacuum decompression is carried out, and the butylacetate comprising mevastatin is separated by continuous print, and concentrated solution is cooled to 0-5 DEG C.Mevastatin crude product is obtained after spending the night.
Experiment F
10L fermented liquid phosphoric acid adjust ph to 3.7, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 13%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 8.5 hours, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
The list of experiment A-F mevastatin crude yield
| Group | Crude product mevastatin yield (%) |
| Experiment A | 78.0 |
| Experiment B | 33.0 |
| Experiment C | 76.5 |
| Experiment D | 70.0 |
| Experiment E | 60.0 |
| Experiment F | 91.2 |
As can be seen from above form crude product mevastatin yield data, the mevastatin crude yield of experiment F reaches 91.2%, far above the mevastatin crude yield of experiment A-E.Experiment A-E is comparative example, and experiment F is the method for the invention, and therefore, the method for the invention is better than the method for comparative example, significantly improves mevastatin crude yield.
embodiment 2
10L fermented liquid phosphoric acid adjust ph, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 8.5 hours, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
| Pretreated pH value | Crude product mevastatin yield (%) |
| 2.0-2.5 | 82.0 |
| 2.5-3.0 | 85.0 |
| 3.0-3.5 | 86.0 |
| 3.5-4.0 | 90.0 |
| 4.0-4.5 | 85.0 |
| 4.5-5.0 | 82.0 |
| 5.0-5.5 | 80.0 |
| 5.5-6.0 | 80.0 |
After receiving fermented liquid, carry out acidizing pretreatment, as can be seen from upper table experimental result, pre-treatment pH value has significant impact for mevastatin crude yield, and when pH value is 3.0-4.0, corresponding mevastatin crude yield is the highest.
embodiment 3
10L fermented liquid phosphoric acid adjust ph to 2.3, filter the bacterium slag obtained and dry up, filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 8.5 hours, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
| Bacterium slag water content | Crude product mevastatin yield (%) |
| 5％ | 84.8 |
| 10％ | 85.0 |
| 20％ | 84.8 |
| 30％ | 83.8 |
| 40％ | 82.0 |
| 50％ | 79.2 |
| 60％ | 76.4 |
| 76％ | 71.8 |
As can be seen from the crude product mevastatin yield of above form, the water content of bacterium slag has significant impact to crude product mevastatin yield, when water content reduces to 40%, this impact starts to diminish, and therefore water content can not as preferred experimental result higher than 40%.
embodiment 4
10L fermented liquid phosphoric acid adjust ph to 3.7, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, cyclisation institute spended time is 7-9 hour, filters, with toluene wash filter residue 3 times, and washings is mixed with filtrate and leaves standstill, complete extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the organic solvent A of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking by organic solvent A, dries thick wet product for about 50 DEG C.
| Organic solvent A | Crude product mevastatin yield (%) |
| Normal hexane | 88.0 |
| Acetone | 78.0 |
| Ethanol | 80.0 |
| Butyl ester | 80.0 |
| Propyl carbinol | 79.0 |
As can be seen from the crude product mevastatin yield of above form, in this step, the selection of organic solvent has significant impact to crude product mevastatin yield, and wherein normal hexane is optimum selection.
embodiment 5
10L fermented liquid phosphoric acid adjust ph to 2.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 6
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 7
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 8
10L fermented liquid phosphoric acid adjust ph to 6.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 9
10L fermented liquid phosphoric acid adjust ph to 3.4, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 10
10L fermented liquid phosphoric acid adjust ph to 5.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 32%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 11
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 5%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 12
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 5%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 13
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 5%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 14
10L fermented liquid phosphoric acid adjust ph to 5.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 5%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 15
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 76%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 16
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 76%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 17
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 76%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 18
10L fermented liquid phosphoric acid adjust ph to 5.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 76%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 19
10L fermented liquid phosphoric acid adjust ph to 2.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 20
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 21
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 22
10L fermented liquid phosphoric acid adjust ph to 6.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 23
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 24
10L fermented liquid phosphoric acid adjust ph to 5.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 25
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 60%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 26
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 60%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 27
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 60%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 28
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 29
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 30
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the normal hexane of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with normal hexane, dries thick wet product for about 50 DEG C.
embodiment 31
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 32
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 33
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 34
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 35
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 36
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 37
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 38
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 39
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the butyl ester of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with butyl ester, dries thick wet product for about 50 DEG C.
embodiment 40
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 41
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 42
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 43
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 44
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 45
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 46
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 47
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 48
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the ethanol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains with after alcohol immersion, dries thick wet product for about 50 DEG C.
embodiment 49
10L fermented liquid phosphoric acid adjust ph to 4.5, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 20%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the propyl carbinol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with propyl carbinol, dries thick wet product for about 50 DEG C.
embodiment 50
10L fermented liquid phosphoric acid adjust ph to 3.6, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 50%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the propyl carbinol of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with propyl carbinol, dries thick wet product for about 50 DEG C.
embodiment 51
10L fermented liquid phosphoric acid adjust ph to 3.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 30%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the acetone of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with acetone, dries thick wet product for about 50 DEG C.
embodiment 52
10L fermented liquid phosphoric acid adjust ph to 5.0, filtering the water content that the bacterium slag that obtains dries up to bacterium slag is 40%, and filtrate detects laggard row relax.Put in extractor by the bacterium slag dried up and toluene, the ratio of toluene and bacterium slag is 6: 1 (V/W), cyclisation, and cyclisation institute spended time is 7-9 hour, filters, and with toluene wash filter residue 3 times, and is mixed with filtrate by washings and leaves standstill, completing extraction.Dry up waste residue, and the waste liquid of waste residue and extraction is carried out environmental protection treatment.
The distillation of extraction filtrate decompression, reclaims toluene and water, after underpressure distillation terminates, reduces concentrated solution temperature to 15 DEG C, adds the acetone of same temperature, insulated and stirred, crystallization.Filter, filter cake vacuumizes, and drains after soaking with acetone, dries thick wet product for about 50 DEG C.
Claims (4)
1. the method for isolated or purified mevastatin from fermented liquid, is characterized in that the method comprises the following steps:
(1) pre-treatment is carried out in the fermented liquid acid containing mevastatin, filter, bacterium slag dries up;
(2) bacterium slag and extraction solvent drop in extractor, insulation cyclisation, filtration washing;
(3) concentrated, crystallization, filtering drying obtains mevastatin crude product;
Wherein the acidifying fermentation liquid pH value of step (1) pre-treatment gained is 3.0-4.5, and it is 5%-40% that bacterium slag dries up rear water content.
2. method according to claim 1, after it is characterized in that step (3) crystallization, adds the organic solvent that mevastatin solubleness is low.
3. method according to claim 2, is characterized in that one or more that described organic solvent is selected from acetone, butylacetate, normal hexane, ethanol.
4. method according to claim 1, is characterized in that step (3) crystallization is filtered the filter cake obtained and vacuumized, drains, dry thick wet product with organic solvent after soaking.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210281403.9A CN102827123B (en) | 2012-08-02 | 2012-08-02 | Separation and extraction process for Mevastatin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210281403.9A CN102827123B (en) | 2012-08-02 | 2012-08-02 | Separation and extraction process for Mevastatin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102827123A CN102827123A (en) | 2012-12-19 |
| CN102827123B true CN102827123B (en) | 2015-04-22 |
Family
ID=47330461
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210281403.9A Active CN102827123B (en) | 2012-08-02 | 2012-08-02 | Separation and extraction process for Mevastatin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102827123B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN184325B (en) * | 1996-12-06 | 2000-08-05 | Helix Biotech Ltd | |
| WO2001081611A1 (en) * | 2000-04-27 | 2001-11-01 | Biocon India Limited | A novel process for the manufacture and purification of compactin |
| CN1406283A (en) * | 2000-02-24 | 2003-03-26 | 拜奥盖尔药厂有限公司 | Method of purifying a fermentation broth |
| CN1504454A (en) * | 2002-12-05 | 2004-06-16 | 上海天伟生物制药有限公司 | Method for purifying mevastatin (forebody of pravastatin) from microorganism fermentation liquor |
| ES2239533B1 (en) * | 2004-03-01 | 2006-12-16 | Ercros Industrial, S.A. | PROCEDURE FOR OBTAINING COMPACTINE. |
| WO2008059466A1 (en) * | 2006-11-17 | 2008-05-22 | Ranbaxy Laboratories Limited | Production of compactin using new mutant strain of pencillium citrinum atcc 38065 |
| CN102046582A (en) * | 2008-04-02 | 2011-05-04 | 帝斯曼知识产权资产管理有限公司 | Pravastatin extraction |
| CN102086183A (en) * | 2009-12-04 | 2011-06-08 | 北大方正集团有限公司 | Method for extracting and separating statins substance |
| CN102344426A (en) * | 2010-07-30 | 2012-02-08 | 北大方正集团有限公司 | Method for extracting and purifying lovastatin |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20020071635A (en) * | 2001-03-07 | 2002-09-13 | 코바이오텍 (주) | A purification method of pravastatin precursor compactin and derivatives thereof using HP20 resin and acetone |
-
2012
- 2012-08-02 CN CN201210281403.9A patent/CN102827123B/en active Active
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IN184325B (en) * | 1996-12-06 | 2000-08-05 | Helix Biotech Ltd | |
| CN1406283A (en) * | 2000-02-24 | 2003-03-26 | 拜奥盖尔药厂有限公司 | Method of purifying a fermentation broth |
| WO2001081611A1 (en) * | 2000-04-27 | 2001-11-01 | Biocon India Limited | A novel process for the manufacture and purification of compactin |
| CN1504454A (en) * | 2002-12-05 | 2004-06-16 | 上海天伟生物制药有限公司 | Method for purifying mevastatin (forebody of pravastatin) from microorganism fermentation liquor |
| ES2239533B1 (en) * | 2004-03-01 | 2006-12-16 | Ercros Industrial, S.A. | PROCEDURE FOR OBTAINING COMPACTINE. |
| WO2008059466A1 (en) * | 2006-11-17 | 2008-05-22 | Ranbaxy Laboratories Limited | Production of compactin using new mutant strain of pencillium citrinum atcc 38065 |
| CN102046582A (en) * | 2008-04-02 | 2011-05-04 | 帝斯曼知识产权资产管理有限公司 | Pravastatin extraction |
| CN102086183A (en) * | 2009-12-04 | 2011-06-08 | 北大方正集团有限公司 | Method for extracting and separating statins substance |
| CN102344426A (en) * | 2010-07-30 | 2012-02-08 | 北大方正集团有限公司 | Method for extracting and purifying lovastatin |
Non-Patent Citations (6)
| Title |
|---|
| A new producer of mevastatin;褚以文 等;《中国抗生素杂志》;19990228;第24卷(第1期);4-6 * |
| Matilde Manzoni,等.Production and purification of statins from Aspergillus terreus strains.《Biotechnology Techniques》.1998,第12卷(第7期),529-532. * |
| Tippabust Eksangsri,等.Extraction of compactin (ML-236B): equilibrium distribution in water-ethyl acetate system.《Chemical Engineering and Processing》.2004,第43卷(第10期),1203-1210. * |
| Tippabust Eksangsri,等.Membrane-based extraction of cholesterol lowering drug: Effect of membrane type on extraction.《Separation and Purification Technology》.2005,第46卷(第1-2期),63-71. * |
| 美伐他汀提取工艺研究;李敏;《天津大学工程硕士学位论文》;20090415;17 * |
| 美伐他汀提取工艺研究;王健 等;《河北工业科技》;20100131;第27卷(第1期);35-37、40 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102827123A (en) | 2012-12-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0702679B1 (en) | Process for the isolation of lovastatin | |
| US20140309451A1 (en) | Purification of 3-hydroxypropionic acid from crude cell broth and production of acrylamide | |
| JP3502419B2 (en) | Method for producing lactic acid and lactic acid ester | |
| WO1994029292A9 (en) | Process for the isolation of lovastatin | |
| WO2013192451A1 (en) | Dehydration of 3-hydroxypropionic acid to acrylic acid | |
| CN1406283A (en) | Method of purifying a fermentation broth | |
| JP2011177159A (en) | Method for isolation and purification of lactic acid from lactic fermentation liquid | |
| CN1206363C (en) | Process for obtaining of HMG-CO a reductase inhibitors of high purify | |
| US7052886B2 (en) | Process for the isolation of lovastatin | |
| CN108164576B (en) | Method for extracting abamectin | |
| CN102827123B (en) | Separation and extraction process for Mevastatin | |
| CN103896956B (en) | A kind of method extracting sesamin from sesame seed coat | |
| CN102875505A (en) | Process for extracting and refining Mevastatin | |
| CN111423252A (en) | Comprehensive treatment method of rhamnolipid fermentation liquor | |
| US20110201054A1 (en) | Process for improved recovery of fermentation products from intracellular and extracellular presence | |
| CN102633642B (en) | Method for preparing pleuromutilin crystal | |
| CN103224547B (en) | Daptomycin separation and purification method | |
| JP6692229B2 (en) | Method for producing 3HB ester | |
| CN103664848B (en) | A kind of extracting method of mycophenolic acid | |
| CN106008624B (en) | A kind of method for crystallising improving avilamycin active principle A, B content | |
| CN101709072B (en) | Method for efficiently extracting and purifying natamycin | |
| US11795482B2 (en) | Method for preparing organic molecules by anaerobic fermentation | |
| CN101168758A (en) | Method for purifying trichodermisin | |
| CN103772248B (en) | A kind of acry radical donor preparation method of simvastatin side chain | |
| WO2009078033A2 (en) | Isolation and recovery of simvastatin in lactone form or in the form of an acid salt from the harvested fermentation broth |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| EE01 | Entry into force of recordation of patent licensing contract | ||
| EE01 | Entry into force of recordation of patent licensing contract |
Application publication date: 20121219 Assignee: Guangdong Blue Treasure Pharmaceutical Co., Ltd. Assignor: Xinbeijiang Pharmaceutical Co., Ltd., Lizhu Group Contract record no.: 2018440000106 Denomination of invention: Separation and extraction process for Mevastatin Granted publication date: 20150422 License type: Common License Record date: 20180816 |