CN106117071B - A kind of membrane filtration isolation and purification method of levodopa conversion fluid - Google Patents
A kind of membrane filtration isolation and purification method of levodopa conversion fluid Download PDFInfo
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Abstract
The invention discloses a kind of membrane filtration isolation and purification methods of levodopa conversion fluid, this method obtains the conversion fluid containing levodopa by tyrosine phenol lyase bioconversion, the pH of conversion fluid is adjusted to highly acid with acid solution, after appropriate heating clear liquid is collected using ceramic membrane filter system, obtained ceramic membrane filter clear liquid is collected, adds in stirring heating decoloration after activated carbon, alkali tune crystallizes after crossing cleaner liquid, it refilters and is dried after being crystallized, obtain levodopa anhydrous crystal powder.This method is on the basis of industrialized production is realized, remaining substrate, albumen and the pigment in conversion process have been effectively removed, has produced the anhydrous crystal powder properties height of acquisition, light transmittance is high, total impurities is by HPLC appearances area percentage no more than 0.1%, and the crystallinity of crystalline powder is more than 90%.
Description
Technical field
The present invention relates to biochemical industry and field is isolated and purified, relates in particular to a kind of film mistake of levodopa conversion fluid
Filter isolation and purification method.
Background technology
Levodopa (3,4-dihydroxylphenylalanine, Levodopa), also known as L-3,4 dihydroxyphenylalanine, chemical name are
3,4- Dihydroxyphenylalanines, CAS：59-92-7；White crystalline powder, odorless, tasteless；It is a kind of important in organism
Bioactive substance is the important intermediate during l-tyrosine to catechol or the biochemical metabolism approach of melanin.Its
Structural formula is as follows：
Levodopa and Benserazide (such as Madopar) treatment common geriatric disease Parkinson's disease have more than 40 years
History is still to treat the most effective drug of Parkinson's disease, and better tolerance so far, can not only improve bradykinesia, alleviates main
Symptom, and the service life of Parkinsonian can be extended.Levodopa ranking 100,2009 in world's best-selling drugs in 2007
Realize that global yield reaches 250 tons, consumption sum in 2010 is more than 1,500,000,000 dollars.Global yield in 2015 close to 1000 tons,
Sales volume is more than more than 30 hundred million U.S. dollars.
Domestic and international scientist is being continually striving to improve the production yields of levodopa.It is reported that the production of levodopa
Method has extracted form natural plant method, chemical synthesis and microbial enzyme conversion method.
Traditional preparation method is to be extracted from plant cat beans through science, and 103641730 A of Chinese patent CN are disclosed
A kind of preparation method of levodopa flattens cat beans by machinery, and with acetic acid flooding, leaching liquor removes egg with trichloroacetic acid
In vain, cream is condensed into, with water dissolution, pH is adjusted to isoelectric point precipitation coarse crystallization, with hot acid water dissolution, activated carbon decolorizing, room temperature weight
Crystallization.103664669 A of Chinese patent CN disclose a kind of process for preparing high-purity levodopa, using cat beans as original
Material, is extracted, extracting solution is enriched with levodopa and is separated by electrodialysis with sour water.Plant extraction method is due to being subject to original
Expect the limitation in source, yield is small, far from meeting current market demand.
The chemical synthesis of levodopa is more.At present, the chemical method of industrialized production levodopa bulk pharmaceutical chemicals comes from
1987, vanillin (vanillic aldehyde) and hydantoin (hydantoins) were, it is necessary to which the reaction by 8 steps is made.But run into one
A main difficulty is the mixture that the product being synthesized is D- types and L-type, and the process control difficulties that the two is separated are big,
Exist simultaneously the problems such as production cost is high, environmental pollution is serious.
Using micro-organisms levodopa, some enzymes in metabolic pathway are exactly make use of, such as tyrosinase, tyrosine
Different substrates is transformed into levodopa by phenols cracking enzyme, transferase, can obtain substantial amounts of levodopa, it is given birth to by many countries
Production has carried out substantial amounts of research, takes many different synthetic methods.The end of the sixties in last century, some external scholars start to cause
Power is in the research of microbial enzyme method synthesis levodopa.The beginning of the seventies, Japanese scholars Yamada etc. synthesizes microbial enzyme method left
Rotation DOPA conducts in-depth research, and work was put into aginomoto company in 1993 by the optimization of strain improvement and fermentation condition
Industry metaplasia is produced.103122361 A of Chinese patent CN are disclosed by building recombinant strain, provide one kind with l-tyrosine
The method that levodopa is synthesized for substrate.104726513 A of Chinese patent CN disclose the side that a kind of enzyme process prepares levodopa
Method produces tyrosinase with Pseudomonas Maltophilia (Pseudomonas maltophilia), is converted by raw material of l-tyrosine
Prepare levodopa.However, preparing levodopa currently with bioanalysis realizes that industrialized report is actually rare.Existing life
Object method prepares that levodopa is long there are the reaction time, and by-product is more, and conversion ratio is low, and substrate residual concentration is high, finished product color and luster compared with
A series of problems, such as difference, crystallinity is relatively low, and impurity content is high.
The content of the invention
Goal of the invention：It is filtered, suitable for industrialized production based on ceramic membranous system the object of the present invention is to provide a kind of
The membrane filtration isolation and purification method of levodopa crystal powder, for obtain in high yield, high-purity, high-crystallinity, high transparency
Levodopa anhydrous crystal powder.
Technical solution：In order to realize foregoing invention purpose, a kind of membrane filtration separation of levodopa conversion fluid of the invention
Purification process, by tyrosine phenol lyase bioconversion obtain containing levodopa, tyrosine phenol lyase, remaining substrate,
Albumen, pigment, the levodopa conversion fluid of salt, levodopa content, will with acid solution not less than 100~120g/L in conversion fluid
The pH of conversion fluid is adjusted to highly acid, clear liquid is collected using ceramic membrane filter system after appropriate heating, until levodopa in trapped fluid
When concentration is down to 5~8g/L, the ceramic membrane filter clear liquid collected adds in stirring heating decoloration after activated carbon, after crossing cleaner liquid
Alkali tune crystallizes, and refilters and is dried after being crystallized, and obtains levodopa anhydrous crystal powder.
The present invention is that the conversion fluid obtained based on the reaction of tyrosine phenol lyase enzyme process carries out levodopa anhydrous crystal powder
The separation and purifying at end.The tyrosine phenol lyase enzyme process is in coenzyme (pyridoxime 5-phosphate) using tyrosine phenol lyase
It under the conditions of existing, is stirred with basic substrate, reaction generates levodopa.Basic substrate is catechol/pyruvic acid ammonium
Or catechol/pyruvic acid/concentrated ammonia liquor.
As shown in formula (I), tyrosine phenol lyase is in NH4 +It, can be by the catechol of a molecule under conditions of being present in excess
Pyruvic acid with a molecule is through single step reaction biosynthesis levodopa.The residue of the reaction includes：Extra pyruvic acid, neighbour
The pigments such as the impurity such as benzenediol, ammonium salt, zymoprotein and catechol oxide, residual enzyme protein solution.
Ceramic membrane of the present invention is microfiltration membranes made of conventional ceramic membrane material, and ceramic membrane element aperture is 50-
200nm, be preferably 50nm, ceramic membrane material is preferably zirconium dioxide, single film be 19 passages, proof pressure 0-0.6Mpa.
It must be it is noted that ceramic membrane materials include but not limited to the materials such as aluminium oxide, titanium oxide, zirconium oxide, preferably membrane component material
For zirconium dioxide, chemical property is stablized, and acidproof, alkaline-resisting, organic solvent-resistant, mechanical strength is big, can rinse repeatedly；It is antimicrobial
Ability is strong；High temperature resistant；Filtering accuracy is high, and separative efficiency is good, and service life is long.Ceramic membrane of the present invention is existing production
Product, but it is very outstanding to the filter effect of conversion fluid of the levodopa content not less than 100~120g/L.
The conversion fluid tune acid pH is adjusted to highly acid, i.e., 0.95~1.05, and conversion fluid temperature rises to 30~40 DEG C when adjusting acid；
Acid solution used includes but not limited to the combination of any one or more in hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, formic acid, acetic acid.It adjusts
Sour purpose is all to dissolve the levodopa crystal in conversion fluid, while zymoprotein can accelerate denaturation to flocculate with this condition.
Between 30~40 DEG C, pH is adjusted to 3.9~4.3, is slowly stirred 1~2 for the conversion fluid alkali tune crystallization temperature control
Hour after the generation of a large amount of crystal, is cooled to 5~10 DEG C, crystal is collected by filtration after standing；Lye used in alkali tune crystallization includes hydrogen
The combination of any one or more in sodium hydroxide solution, potassium hydroxide solution, ammonia spirit.
As the further optimization of the present invention, in order to improve the primary crystallization quality of finished product and yield, cost, alkali tune are saved
During crystallization filtering, with uniformly spray 2~3 times of a small amount of pure water, crystal is collected after drying finally foam washing with pure water once.
Ceramic membrane filter clear liquid is added in 1.0~2.0% activated carbon by quality percent by volume, is warming up to 50~55 DEG C,
Keep the temperature 2 it is small when and be slowly stirred.Pigment in clear liquid is tightly held by activated carbon, wherein containing minimal amount of zymoprotein in acid condition
Under be denatured flocculation, further improve separative efficiency.
As the further optimization of the present invention, antioxidant is additionally added before alkali tune crystallization, the concentration in total feed liquid is
0.5~1.0g/L, the antioxidant includes NaHSO3、Na2SO3、Na2S2O5Or its sylvite, vitamin C or its sodium salt or potassium
The combination of any one or more in salt.
Drying steps after alkali tune crystallization are that 5 ± 0.5cm of solid accumulation is thick, and under temperature 45 C, microwave power control exists
7 ± 0.5kW, vacuum degree -0.099~-0.092Mpa, dry 20~30min.
Explanation is needed exist for, tyrosine phenol lyase Biocatalysis method of the present invention is known technology, used
Tyrosine phenol lyase can directly buy acquisition, and the known restructuring E.coli fermentation tables containing tyrosine phenol lyase can also be used
Up to obtaining the thalline zymotic fluid containing enzyme, then concentrate through film, mechanical breaking-wall method, bactofugation slag obtain concentration enzyme solution.
The levodopa anhydrous crystal powder yield that the method for the invention obtains is not less than 85%.What is obtained is left-handed more
Bar anhydrous crystal powder, by HPLC appearances area percentage no more than 0.1%, the crystallinity of anhydrous crystal is total impurities
More than 90%, crystalline powder grain diameter size is 120 μm~250 μm.
Advantageous effect：The method of the present invention, using ceramic membrane filter system, not only improves on the basis of realization is industrialized
The yield of product, while production efficiency is substantially increased, shorten production cycle, the production cost of reduction so that commercialization
Production has more scale.Using vacuum microwave drying technology, reduce the effect of the external force such as the drum hot wind by traditional drying, stirring and make
Into product loss, avoid levodopa through high-temperature oxydation destroy or decompose after caused product appearance brown stain, improve product do
Moisture homogeneity after dry；The crystalline powder quality that this method obtains is substantially better than EP and USP requirements, and total impurities presses HPLC appearances
Area percentage is not more than 0.1% (EP 1.0%, USP 1.1%), and single impurity presses HPLC appearance area percentages
No more than 0.1% (EP and USP highests are 0.5%), the crystallinity of crystalline powder is more than 90%.
Description of the drawings
Fig. 1 is the efficient liquid phase chromatographic analysis result of the embodiment of the present invention 2；
Fig. 2 is the efficient liquid phase chromatographic analysis result of the embodiment of the present invention 3；
Fig. 3 is levodopa and the (conversion of the HPLC collection of illustrative plates of catechol situation of change in 1 conversion process of the embodiment of the present invention
Carry out to 1 it is small when 40 minutes)；
Fig. 4 is levodopa and the (conversion of the HPLC collection of illustrative plates of catechol situation of change in 1 conversion process of the embodiment of the present invention
Carry out to 3 it is small when)；
Fig. 5 is levodopa and the (conversion of the HPLC collection of illustrative plates of catechol situation of change in 1 conversion process of the embodiment of the present invention
Carry out to 4.5 it is small when).
Specific embodiment
The present invention is further described With reference to embodiment.
1 tyrosine phenol lyase living things catalysis technique of embodiment
Xiang Shuizhong launches initial acetone acid, and pH is controlled 7.0~9.0 with concentrated ammonia liquor；Add antioxidant, chelating successively again
Agent, ammonium salt, pyridoxime 5-phosphate are again controlled pH 7.0~9.0 with concentrated ammonia liquor, add in the weight that concentration is 80~200g/L
The enzyme solution of group tyrosine phenol lyase is passed through nitrogen, adjustment temperature to reaction temperature, and ammonium hydroxide adjusts pH to 7.0~9.0, adds in
Catechol starts to react；It is 1: 1~1.5 that initial basis substrate catechol launches molar ratio with pyruvic acid and its sodium salt.It
Afterwards, substrate catechol and pyruvic acid (or pyruvic acid ammonium, hereafter unless otherwise indicated equal pyruvic acid) are added respectively per 10min
For 2g/L, catechol it is final add concentration be 50~70g/L (being calculated using initial volume), pyruvic acid finally add concentration as
50~70g/L (is calculated) with initial volume, as substrate catechol residual >=5g/L, stops feeding intake, continue stir 0.5h~
Reaction terminates after 1h, obtains levodopa conversion fluid.The 95% of reaction conversion ratio, levodopa content is up to 100 in conversion fluid
~120g/L.
In step of converting, it is also necessary to add the ammonium salt of 0.2mol/L~0.3mol/L, include but not limited to ammonium formate, acetic acid
Any one or more combination in ammonium, ammonium chloride, ammonium sulfate.
After putting into pyruvic acid or its ammonium salt, also put into the antioxidant of substrate quality 1%~5%, antioxidant include but
It is not limited to NaHSO3、Na2SO3、Na2S2O5, vitamin C and its sodium salt the combination of any one or more.
After putting into pyruvic acid and antioxidant, the chelating agent of 0.5g/L~1.5g/L is also put into, chelating agent includes but unlimited
In any one of tartaric acid and its k-na salt, ethylenediamine tetra-acetic acid (EDTA) and its disodium salt.
As shown in Figures 3 to 5, catalytic reaction carried out 4.5 it is small when, levodopa content gradually increases in system, adjacent benzene
Diphenol concentration always≤5g/L；At the end of reaction, the remaining content of catechol only has 0.71g/L.
2 2500L transformation systems of embodiment
Levodopa conversion fluid 2500L, the concentration 110g/L that Example 1 obtains add in concentrated hydrochloric acid tune pH to 0.96, dense
Hydrochloric acid dosage is 190kg.Feed temperature rises to 30~40 DEG C, opens ceramic membranous system filtering.Select Jiangsu my high-tech stock long
The ceramic membranous system of part Co., Ltd, ceramic membrane element aperture are 50nm, and material is zirconium dioxide, and single film is 19 passages, film
The gross area is 21.2 square metres.
Clear liquid is collected using ceramic membrane, when reaching minimum internal circulating load, about 500L systems start stream plus purified water.Detection
The content of levodopa in trapped fluid when being down to 5g/L, stops filtering, and trapped fluid total volume is 530L.Collection obtains clear
Liquid product is 3000L, and the content of levodopa is 89g/L in clear liquid, therefore the yield of ceramic membrane is 97.1%.
Into the 3000L levodopa clear liquids of collection add in 36kg activated carbon, be warming up to 52 DEG C, be slowly stirred 2 it is small when.
After heat preservation, this material removes activated carbon through press filtration cylinder, and clear liquid is pressed into essence after accurate filter and dries warding off in hired car
In porcelain reaction kettle.Stirring is opened, the sodium sulfite solution 14L that concentration is 150g/L is pressed into reaction kettle, until total feed liquid moderate resistance
Oxidant concentration is 0.5~1.0g/L.
35 DEG C or so of control material temperature opens stirring, is slowly added to 30% NaOH solution, pH is adjusted to 3.9, slowly
Stir 2 it is small when, start to cool down；When temperature is down to 8 DEG C, stop stirring, stand still for crystals 5 it is small when.
Crystal is collected by centrifugation with basket centrifuge.During centrifugation, with water spray is purified 2 times, each dosage is 6L water, is finally used
Bubble is washed once, and dosage is 10L water.It is 290kg that Total crystal weight is collected after drying, is packed into layer of polyethylene bag, to be dried.
To after centrifugation crystal material carry out vacuum microwave drying, dried material 5 ± 0.5cm of thickness, 45 DEG C of drying temperature,
Microwave power control is in 7 ± 0.5kW, vacuum degree -0.099~-0.092Mpa, 20~30min of dry duration.Total weight after drying
For 245.6kg, therefore primary crystallization yield is 91.0%, total recovery 89.3%.
After testing, the product dried meets EP and USP requirements：Total impurities presses HPLC appearances area percentage not
More than 0.1%, for the crystallinity of crystalline powder more than 90%, grain diameter is 120 μm~250 μm.
By EP requirements, each index is respectively：Appearance is white crystalline powder, differentiates that IR is consistent with reference substance IR collection of illustrative plates,
Sample size measure main peak retention time is consistent with reference substance peak retention time, and it is molten that solution colour is not deeper than BY6 standard colorimetrics
Liquid, loss on drying 0.19%, pH 5.7, residue≤0.1%, heavy metal≤10mg/kg, related substance is qualified, optical purity
For 99.8%, content is detected as 99.3% with titration.
By USP requirements, indices are respectively：Appearance is white crystalline powder, differentiates IR and reference substance IR collection of illustrative plates one
Cause, sample size measure main peak retention time it is consistent with reference substance peak retention time, loss on drying 0.19%, optical activity be-
165 °, residue≤0.1%, heavy metal≤10mg/kg, content is detected as 99.1% with high performance liquid chromatography, defects inspecting symbol
Close requirement.
3 5000L transformation systems of embodiment
Levodopa conversion fluid 5000L, the concentration 115g/L that Example 1 obtains add in concentrated nitric acid tune pH to 1.04, dense
Nitric acid dosage is 325kg, and feed temperature is risen between 30-40 DEG C, opens ceramic membranous system filtering.Select Jiangsu my high-tech long
The ceramic membranous system of limited company, ceramic membrane element aperture are 50nm, and material is zirconium dioxide, and single film is 19 passages,
The film gross area is 21.2 square metres.
Clear liquid is collected using ceramic membrane, when reaching minimum internal circulating load, about 700L systems start stream plus drinking water.Detection
The content of levodopa in trapped fluid when being down to 8g/L, stops filtering, and trapped fluid total volume is 830L.The clear liquid being collected into
Volume is 6200L, and the content of levodopa is 90.6g/L in clear liquid, therefore the yield of ceramic membrane is 97.7%.
Into the 6200L levodopa clear liquids of collection add in 62kg activated carbon, be warming up to 53 DEG C, be slowly stirred 2 it is small when.
After heat preservation, this material removes activated carbon through press filtration cylinder, and clear liquid is pressed into essence after accurate filter and dries warding off in hired car
In porcelain reaction kettle.Stirring is opened, the vitamin c solution 25L that concentration is 150g/L is pressed into reaction kettle, until total feed liquid moderate resistance oxygen
Agent concentration is 0.5~1.0g/L.
35 DEG C or so of control material temperature opens stirring, is slowly added to 30% potassium hydroxide solution, pH is adjusted to 4.3,
Be slowly stirred 2 it is small when after, start to cool down；When temperature is down to 7 DEG C, stop stirring, stand still for crystals 5 it is small when.
Crystal is collected by centrifugation with basket centrifuge.During centrifugation, with water spray is purified 2 times, each dosage is 6L water, is finally used
Bubble is washed once, and dosage is 10L water.It is 594kg that Total crystal weight is collected after drying, is packed into layer of polyethylene bag, to be dried.
To after centrifugation crystal material carry out vacuum microwave drying, dried material 5 ± 0.5cm of thickness, 45 DEG C of drying temperature,
Microwave power control is in 7 ± 0.5kW, vacuum degree -0.099~-0.092Mpa, 20~30min of dry duration.Total weight after drying
For 506kg, therefore primary crystallization yield is 90.1%, total recovery 88.0%.
After testing, the product dried meets EP and USP requirements.Total impurities presses HPLC appearances area percentage not
More than 0.1%, for the crystallinity of crystalline powder more than 90%, grain diameter is 120 μm~250 μm.
By EP requirements, each index is respectively：Appearance is white crystalline powder, differentiates that IR is consistent with reference substance IR collection of illustrative plates,
Sample size measure main peak retention time is consistent with reference substance peak retention time, and it is molten that solution colour is not deeper than BY6 standard colorimetrics
Liquid, loss on drying 0.16%, pH 5.5, residue≤0.1%, heavy metal≤10mg/kg, related substance is qualified, optical purity
For 99.6%, content is detected as 99.3% with titration.
By USP requirements, indices are respectively：Appearance is white crystalline powder, differentiates IR and reference substance IR collection of illustrative plates one
Cause, sample size measure main peak retention time it is consistent with reference substance peak retention time, loss on drying 0.16%, optical activity be-
164 °, residue≤0.1%, heavy metal≤10mg/kg, content is detected as 99.0% with high performance liquid chromatography, defects inspecting symbol
Close requirement.
Test example
HPLC analyzes the impurity of 3 product of above-described embodiment 2 and embodiment, and result is as shown in Figure 1 and Figure 2.
1 embodiment of table, 2 levodopa anhydrous crystal powder HPLC analysis results
| Retention time | Area | % areas | Highly | Integral type | |
| 1 | 3.523 | 7573 | 0.03 | 928 | VV |
| 2 | 6.298 | 29111395 | 99.96 | 2653746 | BB |
| 3 | 24.105 | 2124 | 0.01 | 256 | BV |
2 embodiment of table, 3 levodopa anhydrous crystal powder HPLC analysis results
| Retention time | Area | % areas | Highly | Integral type | |
| 1 | 3.080 | 9482 | 0.03 | 614 | VV |
| 2 | 4.086 | 29122425 | 99.94 | 2652535 | BB |
| 3 | 25.115 | 9472 | 0.03 | 731 | VB |
Table 1, table 2 correspond to the HPLC collection of illustrative plates of Figure of description Fig. 1, Fig. 2 respectively.The present invention prepares work according to existing production
Skill, used raw material is soluble easily in water, therefore organic solvent need not be used in production process.Impurity content is remote in obtained product
Far below the product that other technique productions obtain, purity is above 99.9%.
The above is only the preferred embodiment of the present invention, it should be pointed out that：For the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (8)
1. a kind of membrane filtration isolation and purification method of levodopa conversion fluid, is contained by tyrosine phenol lyase bioconversion
There is a levodopa conversion fluid of levodopa, tyrosine phenol lyase, remaining substrate, albumen, pigment, salt, it is left in conversion fluid
It revolves DOPA content and is not less than 100 ~ 120g/L, it is characterised in that：
The pH of conversion fluid is adjusted to highly acid with acid solution, clear liquid is collected using ceramic membrane filter system after appropriate heating, until retention
Levodopa concentration is down to 5-8g/L in liquid, and the ceramic membrane filter clear liquid collected adds in stirring heating decoloration after activated carbon,
Alkali tune crystallizes after crossing cleaner liquid, refilters and is dried after being crystallized, and obtains levodopa anhydrous crystal powder；
Between 30 ~ 40 DEG C, pH is adjusted to 3.9 ~ 4.3 for conversion fluid alkali tune crystallization temperature control, be slowly stirred 1 ~ 2 it is small when treat it is big
After measuring crystal generation, 5 ~ 10 DEG C are cooled to, crystal is collected by filtration after standing；
The levodopa anhydrous crystal powder yield that this method obtains is not less than 85%.
2. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 1, it is characterised in that：Institute
Ceramic membrane element aperture in ceramic membrane filter system is stated as 50nm, single film is 19 passages, and material is zirconium dioxide, ceramic membrane
The system gross area is 21.2 square metres.
3. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 1, it is characterised in that：Institute
It states conversion fluid tune acid pH and is adjusted to 0.95 ~ 1.05, conversion fluid temperature rises to 30 ~ 40 DEG C；Acid solution used include hydrochloric acid, sulfuric acid, phosphoric acid,
The combination of any one or more in nitric acid, formic acid, acetic acid.
4. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 1, it is characterised in that：It adjusts
Lye used in alkali crystallization includes the combination of any one or more in sodium hydroxide solution, potassium hydroxide solution, ammonia spirit.
5. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 4, it is characterised in that：It adjusts
During alkali crystallization filtering, with uniformly spray 2 ~ 3 times of a small amount of pure water, crystal is collected after drying finally foam washing with pure water once.
6. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 1, it is characterised in that：Pottery
Porcelain membrane filtration clear liquid is added in 1.0 ~ 2.0% activated carbon by quality percent by volume, is warming up to 50 ~ 55 DEG C, when heat preservation 2 is small and is delayed
Slow stirring.
7. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 4 or 5, feature exist
In：Antioxidant is additionally added before alkali tune crystallization, making its concentration in total feed liquid, the antioxidant includes for 0.5 ~ 1.0g/L
NaHSO3、Na2SO3、Na2S2O5Or the combination of any one or more in its sylvite, vitamin C or its sodium salt or sylvite.
8. a kind of membrane filtration isolation and purification method of levodopa conversion fluid according to claim 1, it is characterised in that：It adjusts
Drying steps after alkali crystallization are that 5 ± 0.5cm of solid accumulation is thick, and under temperature 45 C, microwave power is controlled in 7 ± 0.5 kW,
The Mpa of vacuum degree -0.099 ~ -0.092, dry 20 ~ 30min.
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| US20050277174A1 (en) * | 2004-03-29 | 2005-12-15 | Hidehiko Kumagai | Mutant tyrosine repressor, a gene encoding the same, and a method for producing L-DOPA |
| CN104402748A (en) * | 2014-12-05 | 2015-03-11 | 南宁知本康业生物技术有限公司 | Microwave-assisted method for extracting levodopa from cat beans |
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| US20050277174A1 (en) * | 2004-03-29 | 2005-12-15 | Hidehiko Kumagai | Mutant tyrosine repressor, a gene encoding the same, and a method for producing L-DOPA |
| CN104402748A (en) * | 2014-12-05 | 2015-03-11 | 南宁知本康业生物技术有限公司 | Microwave-assisted method for extracting levodopa from cat beans |
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