CN104004002B - A kind of method directly being prepared 6-amino-penicillanic acid by penicillin fermentation liquid - Google Patents
A kind of method directly being prepared 6-amino-penicillanic acid by penicillin fermentation liquid Download PDFInfo
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- CN104004002B CN104004002B CN201410208804.0A CN201410208804A CN104004002B CN 104004002 B CN104004002 B CN 104004002B CN 201410208804 A CN201410208804 A CN 201410208804A CN 104004002 B CN104004002 B CN 104004002B
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- C07D499/00—Heterocyclic compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. penicillins, penems; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring
- C07D499/21—Heterocyclic compounds containing 4-thia-1-azabicyclo [3.2.0] heptane ring systems, i.e. compounds containing a ring system of the formula:, e.g. penicillins, penems; Such ring systems being further condensed, e.g. 2,3-condensed with an oxygen-, nitrogen- or sulfur-containing hetero ring with a nitrogen atom directly attached in position 6 and a carbon atom having three bonds to hetero atoms with at the most one bond to halogen, e.g. an ester or nitrile radical, directly attached in position 2
- C07D499/42—Compounds with a free primary amino radical attached in position 6
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- C12P37/00—Preparation of compounds having a 4-thia-1-azabicyclo [3.2.0] heptane ring system, e.g. penicillin
- C12P37/06—Preparation of compounds having a 4-thia-1-azabicyclo [3.2.0] heptane ring system, e.g. penicillin by desacylation of the substituent in the 6 position
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Abstract
The invention belongs to biomedicine field, relate to a kind of method directly being prepared 6 aminopenicillanic acids by penicillin fermentation liquid.It is big to there is environmental pollution in the preparation method of existing 6 aminopenicillanic acids, energy consumption consumes height, the technical deficiencies such as product yield is low, in order to overcome above-mentioned technical deficiency, the present invention provides a kind of method directly being prepared 6 aminopenicillanic acids by penicillin fermentation liquid, this method avoid the use of the organic solvent such as butyl acetate and n-butyl alcohol, improve the yield of product, reduce energy expenditure in preparation process simultaneously, it is 6 aminopenicillanic acid production methods of a kind of green energy conservation, is especially suitable for industrially popularization and application.
Description
Technical field
The invention belongs to biomedicine field, be specifically related to one and directly made by penicillin fermentation liquid
The method of standby 6-amino-penicillanic acid.
Background technology
6-amino-penicillanic acid (6-aminopenicillanic acid, 6-APA) is white plates
Crystallization, is the important intermediate synthesizing various semisynthetic penicillins, has many uses.As former
Material carries out modifying for chemical structure, connects the side chain of different structure, it is possible to produce resistance to penicillin
Medicine bacterium is sensitive, the wider array of new semisynthetic penicillin of antimicrobial spectrum, such as ampicillin, amoxycillin
Penicillin, penicillin Vl phenoxymethylpenicillin, and other have the various semi-synthetic of wider antimicrobial spectrum
Penicillin.The annual production of current domestic 6-APA is more than 30000 tons.
Current industrial removal penicillin side chain cleavage becomes 6-APA, mainly has micro bioenzyme catalysis
Cracking process and chemical cleavage method.Along with developing rapidly of biotechnology, use immobilized enzyme
Or immobilized cell produces 6-APA, not only technique is greatly simplified, and economic benefit is obvious, and can
Obtain the 6-APA that purity is higher.Enzyme process is increasingly becoming industrial production 6-APA's in recent years
Main flow.
Chemical cleavage method: the process route for industrial chemical cracking is: extremely low
At a temperature of, first the carboxyl of penicillin is transformed into estersil and protects, then make the acyl on side chain
Amine activates, by forming penicillin substituted imine ether derivant, then under conditions of extremely gentleness,
Optionally hydrolysis chain rupture becomes 6-APA.
Catalyzed by biological enzyme: antibacterial, actinomycetes, yeast and higher fungus are all in nature
PA ase can be produced.Along with the development of modern biotechnology, enzyme strain improvement,
Fermentation automatization, the large-scale of enzyme, enzyme immobilizatio, reactor design, subsequent technique etc.
Every field makes progress simultaneously, and PA ase becomes for the preparation of 6APA the most very much
Ripe.Immobilized enzyme can be reused, and easily separates from reactant liquor, and it is right effectively to prevent
The protein contamination of product and microorganism pollution etc..A certain amount of immobilization is added in reactor
Enzyme, certain density penicillin solution and immobilized enzyme under the effect of stirring, makes enzyme and green grass or young crops
Mycin is fully contacted.Under the catalysis of enzyme, penicillin is constantly cracked into 6-APA and benzene second
Acid, the pH value of solution declines, adds certain density ammonia and make pH value maintain 8.0, when
PH value is not declining and is maintaining 10 minutes, arrives reaction end.Above-mentioned lysate adds
Certain methanol, adjusts pH value to 4.2 with hydrochloric acid, makes 6-APA crystallization, then filter,
It is dried to obtain finished product.
Straight-through technological process: existing technique is that potassium salt of penicillin finished product is dissolved into aqueous solution,
Then crack.And potassium salt of penicillin extracts from aqueous solution and crystallizes out.For contracting further
Short processes flow process, reduces cost and energy consumption, no longer makes penicillium crystallization out.Send out at penicillin
The pre-treating technology of ferment liquid is made suitable process modification, obtains finite concentration and degree of purity
RB liquid (aqueous solution during penicillin extraction) cracks.And lysate is extracted,
Then crystallization obtains 6-APA.This greatly simplifies technological process.
Three of the above technique, is as the development of biotechnology and the continuous of penicillin extraction process
The result of the technological progresses such as improvement.Wherein, chemical method reaction condition requires strict, needs to make
With the chemical reagent of multiple costliness, and require to react at a temperature of extremely low at-40 DEG C.The opposing party
Face produces the organic wastewater of the intractable high concentration of environmental protection.Therefore chemical method is the most superseded.Biological
Method uses immobilized enzyme cracking, and thousands of times of immobilized enzyme energy Reusability, yield is also high than chemical method
3%～5%, water improves production efficiency the most to a great extent, reduces production cost, and
Alleviate environmental protection pressure.Straight-through Process is on the basis of improving penicillin extraction process, enters
One step simple flow, saves benzylpenicillin potassium salt-pepper noise workshop section, decreases the consumption of steam, reduces
Solvent consumption；On the other hand the discharge capacity of the organic wastewater of high concentration is decreased, the biggest
Environmental protection pressure is alleviated in degree.
The 6-APA preparation method that above-mentioned straight-through technique is initiateed by RB still suffers from more shortcoming:
Production process uses the organic solvent such as butyl acetate, n-butyl alcohol in a large number, environment is worked the mischief,
Process steps is long, relates to the rectification-recovery process of above-mentioned solvent, consumes substantial amounts of steam, cold
But water equal energy source.As Chinese patent application CN101735243A discloses a kind of through type 6-
Aminopenicillanic acid preparation method, it comprises the following steps: a, penicillin fermentation liquid is filtered,
Be acidified, butyl alcohol extraction concentrates, decolour to obtain penicillin butyl ester extracting solution；B, penicillin butyl ester carry
Take liquid through alkaline solution back extraction, obtain penicillin salt aqueous solution；C, under vacuum decompression state,
Penicillin salt aqueous solution pours into de-ester tower continuously, and butyl ester proceeds to gas from benzylpenicillin sodium solution
Phase, the benzylpenicillin sodium solution after de-ester is from discharging depressurized system at the bottom of tower, entering with chiller
Storage tank, standby after cooling；D, will cooling after de-ester penicillin salt aqueous solution inject penicillium sp
After element acylase retort carries out enzymatic conversion, add 6-APA crystal seed, growing the grain, crystallize,
It is dried.Complex steps in this preparation method, requires harshness to condition in preparation process, and energy consumption needs
Ask big, and preparation process employs substantial amounts of organic solvent, cause bigger environmental protection pressure.
Chinese patent application CN102925526A discloses the preparation of a kind of 6-amino-penicillanic acid
Method, it comprises the following steps: a, penicillin fermentation liquid are through Ultra filtration membrane and NF membrane
It is concentrated to give concentrated filtrate；B, concentrated filtrate is placed in retort, adds immobilized penicillin
Acylase 3.5-5MU/m3 concentrated filtrate, carries out conversion reaction, obtains 6-amino-penicillanic acid
Conversional solution；C, described conversional solution obtain 6-amino-penicillanic acid filter after activated carbon decolorizing and filtration
Liquid；D, to c step operation described in filtrate in add crystal seed, growing the grain, add crystallizing agent, knot
Brilliant, filter, wash, be dried.This preparation method is relative to patent application CN101735243A
Decrease the use of organic solvent to a certain extent, but the method is in order to prevent penicillin fermentation liquid
Degrading at normal temperatures, its ultrafiltration and nanofiltration operation use-10 DEG C of coolant to control temperature, even if but
So, substantial portion of penicillin is also had to degrade.Therefore, the technology of this preparation method lacks
Point is relatively low for Membrane Filtration Flux, and energy consumption is relatively big, and the yield of product is low and preparation cost is higher.
Summary of the invention
In order to overcome above-mentioned the deficiencies in the prior art, the present invention provides that a kind of technique is simple, safety
The 6-amino-penicillanic acid that energy-conservation, product yield is high leads directly to its preparation process.Of the present invention
A kind of new new method directly being prepared 6-amino-penicillanic acid by penicillin fermentation liquid, it is to avoid
The use of the organic solvent such as butyl acetate and n-butyl alcohol and the ring that thus organic solvent causes
Environment pollution, improves the yield of product, has saved consumption when organic solvent rectification is reclaimed simultaneously
The energy, is the 6-amino-penicillanic acid production method of a kind of green energy conservation.
For solving above-mentioned technical problem, the present invention is practiced by the following technical solutions:
A kind of method directly being prepared 6-amino-penicillanic acid by penicillin fermentation liquid, it include as
Lower step:
A, in penicillin fermentation liquid, add PA ase carry out cracking reaction, reaction knot
Obtain after bundle comprising 6-amino-penicillanic acid, phenylacetic acid and the mixed liquor of penicillin mycelia；
B, mixed liquor is sequentially passed through microfiltration, ultra-filtration and separation removes penicillin mycelia and macromole
Material, is concentrated to give the concentration mixing of 6-amino-penicillanic acid and phenylacetic acid by solution through nanofiltration
Solution；
C, the concentration mixed solution of 6-amino-penicillanic acid and phenylacetic acid is carried out desolventing technology after,
The phenylacetic acid adsorbing separation using resin to concentrate in mixed solution, obtains 6-amino-penicillanic acid
Aqueous solution；
D, regulation 6-amino-penicillanic acid the pH value of aqueous solution to 6-amino-penicillanic acid
Isoelectric point, IP i.e. can get 6-amino-penicillanic acid crystallization, and filtration washing the most i.e. can get this
Bright described 6-amino-penicillanic acid.The isoelectric point, IP of wherein said 6-amino-penicillanic acid is
4.3。
In the preparation method of 6-amino-penicillanic acid described above, the temperature of described cracking reaction
For 26-38 DEG C, in course of reaction, the pH of fermentation liquid is 7.0-8.5.Required for cracking reaction
PA ase consumption adds according to its activity, uses ammonia regulation pH in course of reaction,
When in reactor in 20min after stopping adding ammonia, pH value keeps constant, reaction terminates.
Response time, conversion ratio should be up to 95-98% typically at 1-3 hour.
In the preparation method of 6-amino-penicillanic acid described above, described microfiltration, ultrafiltration and receive
Filter membrane used in filter is organic membrane, ceramic membrane or metal film, wherein the precision of ultrafilter membrane
Quality and the flux of film according to fermentation liquid select, and preferably molecular cut off is
3000--50000 daltonian film group.The molecular weight that dams of described NF membrane is 100～260
Dalton.In the preparation method of 6-amino-penicillanic acid described above, described 6-amino is blue or green
The mass percent of 6-amino-penicillanic acid in mixed solution that concentrates of mould alkanoic acid and phenylacetic acid is
5%-15%.
In the preparation method of 6-amino-penicillanic acid described above, described desolventing technology decolours
Agent is activated carbon, aluminium sesquioxide or a combination thereof.The system of 6-amino-penicillanic acid described above
In Preparation Method, described resin is hypercrosslinked polymeric resin or macroporous adsorbent resin.Wherein, described
Hypercrosslinked polymeric resin model be the one in AH, NDa-150, described macroporous adsorbent resin
Model be Amberlite XAD-4.
In the preparation method of 6-amino-penicillanic acid described above, described step d includes as follows
Step: the aqueous solution controlling 6-amino-penicillanic acid controls at 10～25 DEG C, uses ammonia to adjust
Joint solution ph, to the isoelectric point, IP of 6-amino-penicillanic acid, obtains the knot of 6-amino-penicillanic acid
Crystalline substance, filters, washs, is dried.
In the preparation method of 6-amino-penicillanic acid described above, it is also possible to include that phenylacetic acid takes off
Adsorption step, wherein said phenylacetic acid De contamination comprise the steps: use add ethanol or
The sodium hydroxide solution washing of acetone is adsorbed with the resin of phenylacetic acid and makes phenylacetic acid De contamination.So
Resin is regenerated, and the sodium phenylacetate simultaneously obtained can be answered as raw material through the most refined
In penicillin fermentation produces.
In the preparation method of 6-amino-penicillanic acid of the present invention, whole process does not use
The organic solvent such as butyl acetate and n-butyl alcohol, thus avoid what the volatilization of these organic solvents caused
Environmental pollution, avoids steam, the cooling water equal energy source power reclaiming organic solvent and consuming simultaneously.
Penicillin fermentation liquid is the most degradable, patent application 201210481273.3 in ultrafiltration and
Nanofiltration operation, can utilize-10 DEG C of coolant to control temperature, and it is relatively low that this results in Membrane Filtration Flux,
Energy consumption is big, and yield is low, and cost is high.And the present invention is by enzymolysis, penicillin is cracked into relatively
Stable 6-APA and PAA, it is possible to accomplish to complete membrane filtration under room temperature operating mode, improves
Membrane flux, improves the yield of product, reduces energy consumption and cost.
Preparation method of the present invention compared with prior art, has a following technical advantage:
The most existing straight-through technique produces the later stage use of 6-APA preparation method and uses second in a large number
The organic solvent such as acid butyl ester, n-butyl alcohol, not only technique very complicated, and there is the biggest ring
Packing pressure, after the present invention removes mycelium and macromole impurity by membrane filtration technique, use etc.
Electricity point method for crystallising obtains target product, and its step is simple, does not produce environmental pollution product.And
It is more than 89% that the purity using the target product of this method acquisition is not less than 98.5% yield, high
The purity of 6-APA produced in existing straight-through technique or yield.
2. patent application 201210481273.3 utilizes-10 DEG C of coolant in ultrafiltration and nanofiltration operation
Control temperature, then use PA ase cracking, the most not only cause production process energy consumption
Consuming big, yield is low, and cost improves, and due to the unstability of penicillin, causes at film
Fermentation liquid produces during filtration a large amount of impurity, and then reduces purity and the yield of 6-APA.
The present invention the most i.e. completes the cracking reaction of penicillin so that the flux of membrane filtration is notable
Promoting, purity and the yield of product significantly improve, and filter process of the present invention is the most at normal temperatures
Carry out, energy-conserving and environment-protective.
3. the present invention use in the mixed liquor of Separation of Benzene acetic acid and 6-APA specific optionally
Exclusive resin absorption phenylacetic acid, it is to avoid solvent extraction process.Retaining of resin absorption phenylacetic acid
Rate more than 80%, absorbed portion impurity simultaneously.It is adsorbed with the resin of phenylacetic acid through simple place
Can complete phenylacetic acid after reason to reclaim and resin regeneration, phenylacetic acid can be reused for the life of penicillin
In product, the resin after regeneration can be reused, thus reduces the fortune of preparation method of the present invention
Row cost.
Accompanying drawing explanation
The preparation technology figure of Fig. 1 6-amino-penicillanic acid of the present invention.
Detailed description of the invention
Fig. 1 is the preparation technology figure of 6-amino-penicillanic acid of the present invention.As it can be seen, its
Preparation technology comprises the steps:
A, in penicillin fermentation liquid, add PA ase carry out cracking reaction, reaction knot
Obtain after bundle comprising 6-amino-penicillanic acid, phenylacetic acid and the mixed liquor of penicillin mycelia；
B, mixed liquor is sequentially passed through microfiltration, ultra-filtration and separation removes penicillin mycelia and macromole
Material, is concentrated to give the concentration mixing of 6-amino-penicillanic acid and phenylacetic acid by solution through nanofiltration
Solution；
C, the concentration mixed solution of 6-amino-penicillanic acid and phenylacetic acid is carried out desolventing technology after,
The phenylacetic acid adsorbing separation using resin to concentrate in mixed solution, obtains 6-amino-penicillanic acid
Aqueous solution；
D, regulation 6-amino-penicillanic acid the pH value of aqueous solution to 6-amino-penicillanic acid
Isoelectric point, IP i.e. can get 6-amino-penicillanic acid crystallization, and filtration washing the most i.e. can get this
Bright described 6-amino-penicillanic acid.
In order to further describe the present invention, further illustrate the present invention below by way of specific embodiment
Technical scheme, but described embodiment limit the present invention never in any form.
The preparation method of embodiment 1 6-amino-penicillanic acid of the present invention
The penicillin fermentation liquid 1000mL that concentration is 100000u/mL is placed on temperature control
In the stirred reactor of 28～30 DEG C, stirring.Treat that the temperature of fermentation liquid reaches 28～30 DEG C
Time, add penicillin and be acylated, crack, and constantly dropping concentration is in cracking process
The ammonia of 10%, maintains about 7.0 by pH value in reactor, after stopping adding ammonia
When in 20min, in reactor, pH value keeps constant, reaction terminates.
Mixed liquor after cracking reaction is sequentially passed through the filter of 10 microns, the mistake of 1 micron
Filter removes the penicillin mycelia of solid.The most again mixed solution is sequentially passed through molecular cut off
It is that 3000-5000 is daltonian for the daltonian ultrafilter of 30000-50000 and molecular cut off
Ultrafilter, removes the impurity such as macro-molecular protein.
By the mixed solution after remove impurity through molecular cut off be 100～260 daltonian nanofiltrations
Device, makes mixed solution be concentrated, and the mass concentration being concentrated into 6-APA is 5%.
Activated carbon, aluminium sesquioxide etc. is utilized to be decoloured by concentrated solution, the mixed liquor after being decoloured.
Utilize the hypercrosslinked polymeric resin AH of specific Selective adsorption by the mixed liquor after decolouring
Phenylacetic acid absorption, through solid-liquid separation, respectively obtain the aqueous solution of 6-APA and be adsorbed with benzene
The resin of acetic acid.
The aqueous solution of 6-APA is transferred in crystallizer, in crystallizer temperature control 10～
12 DEG C, adding ammonia in crystallizer, in gradually regulating crystallizer, solution ph is to 6-APA's
Isoelectric point, IP (PI=4.3), obtains the crystallization of 6-APA, filters, washs, obtains this after drying
Invention product.After testing, in described product, the purity of 6-APA is 98.5%, and yield is 89.7%.
It is adsorbed with the resin of phenylacetic acid, washes through adding the sodium hydroxide solution of ethanol or acetone
Washing, make the phenylacetic acid of absorption be desorbed, resin is regenerated, and makes resin be reused, with
Time obtain sodium phenylacetate, can be applied to penicillin fermentation as raw material raw through the most refined
In product.
The preparation method of embodiment 2 6-amino-penicillanic acid of the present invention
The penicillin fermentation liquid 1000mL that concentration is 100000u/mL is placed on temperature control
In the stirred reactor of 30～32 DEG C, stirring.Treat that the temperature of fermentation liquid reaches 30～32 DEG C
Time, add penicillin and be acylated, crack, and constantly dropping concentration is in cracking process
The ammonia of 5%, maintains about 7.4 by pH value in reactor, after stopping adding ammonia
When in 20min, in reactor, pH value keeps constant, reaction terminates.
Mixed liquor after cracking reaction is sequentially passed through the filter of 10 microns, the mistake of 1 micron
Filter removes the penicillin mycelia of solid.The most again mixed solution is sequentially passed through molecular cut off
It is that 3000-5000 is daltonian for the daltonian ultrafilter of 30000-50000 and molecular cut off
Ultrafilter, removes the impurity such as macro-molecular protein.
By the mixed solution after remove impurity through molecular cut off be 100～260 daltonian nanofiltrations
Device, makes mixed solution be concentrated, and the mass concentration being concentrated into 6-APA is 15%.
Activated carbon, aluminium sesquioxide etc. is utilized to be decoloured by concentrated solution, the mixed liquor after being decoloured.
Utilize the hypercrosslinked polymeric resin NDa-150 of specific Selective adsorption by the mixing after decolouring
Phenylacetic acid absorption in liquid, through solid-liquid separation, respectively obtains aqueous solution and the absorption of 6-APA
There is the resin of phenylacetic acid.
The aqueous solution of 6-APA is transferred in crystallizer, in crystallizer temperature control 15～
18 DEG C, adding ammonia in crystallizer, in gradually regulating crystallizer, solution ph is to 6-APA's
Isoelectric point, IP (PI=4.3), obtains the crystallization of 6-APA, filters, washs, obtains this after drying
Invention product.After testing, in described product, the purity of 6-APA is 99.1%, and yield is 90.4%.
It is adsorbed with the resin of phenylacetic acid, washes through adding the sodium hydroxide solution of ethanol or acetone
Washing, make the phenylacetic acid of absorption be desorbed, resin is regenerated, and makes resin be reused, with
Time obtain sodium phenylacetate, can be applied to penicillin fermentation as raw material raw through the most refined
In product.
The preparation method of embodiment 3 6-amino-penicillanic acid of the present invention
The penicillin fermentation liquid 1000mL that concentration is 100000u/mL is placed on temperature control
In the stirred reactor of 34～36 DEG C, stirring.Treat that the temperature of fermentation liquid reaches 34～36 DEG C
Time, add penicillin and be acylated, crack, and constantly dropping concentration is in cracking process
The ammonia of 8%, maintains about 8.0 by pH value in reactor, after stopping adding ammonia
When in 20min, in reactor, pH value keeps constant, reaction terminates.
Mixed liquor after cracking reaction is sequentially passed through the filter of 10 microns, the mistake of 1 micron
Filter removes the penicillin mycelia of solid.The most again mixed solution is sequentially passed through molecular cut off
It is that 3000-5000 is daltonian for the daltonian ultrafilter of 30000-50000 and molecular cut off
Ultrafilter, removes the impurity such as macro-molecular protein.
By the mixed solution after remove impurity through molecular cut off be 100～260 daltonian nanofiltrations
Device, makes mixed solution be concentrated, and the mass concentration being concentrated into 6-APA is 10%.
Activated carbon, aluminium sesquioxide etc. is utilized to be decoloured by concentrated solution, the mixed liquor after being decoloured.
After the macroporous adsorbent resin Amberlite XAD-4 utilizing specific Selective adsorption will decolour
Mixed liquor in phenylacetic acid absorption, through solid-liquid separation, respectively obtain the aqueous solution of 6-APA
With the resin being adsorbed with phenylacetic acid.
The aqueous solution of 6-APA is transferred in crystallizer, in crystallizer temperature control 20～
23 DEG C, adding ammonia in crystallizer, in gradually regulating crystallizer, solution ph is to 6-APA's
Isoelectric point, IP (PI=4.3), obtains the crystallization of 6-APA, filters, washs, obtains this after drying
Invention product.After testing, in described product, the purity of 6-APA is 98.7%, and yield is 91.5%.
It is adsorbed with the resin of phenylacetic acid, washes through adding the sodium hydroxide solution of ethanol or acetone
Washing, make the phenylacetic acid of absorption be desorbed, resin is regenerated, and makes resin be reused, with
Time obtain sodium phenylacetate, can be applied to penicillin fermentation as raw material raw through the most refined
In product.
The preparation method of embodiment 4 6-amino-penicillanic acid of the present invention
The penicillin fermentation liquid 1000mL that concentration is 100000u/mL is placed on temperature control
In the stirred reactor of 35～38 DEG C, stirring.Treat that the temperature of fermentation liquid reaches 35～38 DEG C
Time, add penicillin and be acylated, crack, and constantly dropping concentration is in cracking process
The ammonia of 6%, maintains about 8.5 by pH value in reactor, after stopping adding ammonia
When in 20min, in reactor, pH value keeps constant, reaction terminates.
Mixed liquor after cracking reaction is sequentially passed through the filter of 10 microns, the mistake of 1 micron
Filter removes the penicillin mycelia of solid.The most again mixed solution is sequentially passed through molecular cut off
It is that 3000-5000 is daltonian for the daltonian ultrafilter of 30000-50000 and molecular cut off
Ultrafilter, removes the impurity such as macro-molecular protein.
By the mixed solution after remove impurity through molecular cut off be 100～260 daltonian nanofiltrations
Device, makes mixed solution be concentrated, and the mass concentration being concentrated into 6-APA is 12%.
Activated carbon, aluminium sesquioxide etc. is utilized to be decoloured by concentrated solution, the mixed liquor after being decoloured.
Utilize the hypercrosslinked polymeric resin AH etc. of specific Selective adsorption by the mixed liquor after decolouring
In phenylacetic acid absorption, through solid-liquid separation, respectively obtain the aqueous solution of 6-APA and be adsorbed with
The resin of phenylacetic acid.
The aqueous solution of 6-APA is transferred in crystallizer, in crystallizer temperature control 18～
20 DEG C, adding ammonia in crystallizer, in gradually regulating crystallizer, solution ph is to 6-APA's
Isoelectric point, IP (PI=4.3), obtains the crystallization of 6-APA, filters, washs, obtains this after drying
Invention product.After testing, in described product, the purity of 6-APA is 98.9%, and yield is 89.3%.
It is adsorbed with the resin of phenylacetic acid, washes through adding the sodium hydroxide solution of ethanol or acetone
Washing, make the phenylacetic acid of absorption be desorbed, resin is regenerated, and makes resin be reused, with
Time obtain sodium phenylacetate, can be applied to penicillin fermentation as raw material raw through the most refined
In product.
Although, the most with a general description of the specific embodiments the present invention has been made in detail
Most description, but on the basis of the present invention, it can be modified or improve, this is right
It is apparent from for those skilled in the art, therefore, at the base without departing from spirit of the present invention
These modifications or improvements on plinth, belong to the scope of protection of the invention.
Claims (11)
1. the method directly being prepared 6-amino-penicillanic acid by penicillin fermentation liquid, it comprises the steps:
A, adding PA ase carry out cracking reaction in penicillin fermentation liquid, reaction obtains comprising 6-amino after terminating
Penicillanic acid, phenylacetic acid and the mixed liquor of penicillin mycelia；
B, mixed liquor is sequentially passed through microfiltration, ultra-filtration and separation removes penicillin mycelia and macromolecular substances, by solution through receiving
Filter is concentrated to give the concentration mixed solution of 6-amino-penicillanic acid and phenylacetic acid；
C, the concentration mixed solution of 6-amino-penicillanic acid and phenylacetic acid is carried out desolventing technology after, use resin by mixed for concentrations
Close the phenylacetic acid adsorbing separation in solution, obtain the aqueous solution of 6-amino-penicillanic acid；
D, pH value to the isoelectric point, IP of 6-amino-penicillanic acid of aqueous solution of regulation 6-amino-penicillanic acid i.e. can get 6-amino
Penicillanic acid crystallizes, and filtration washing the most i.e. can get 6-amino-penicillanic acid of the present invention；
Wherein said step d comprises the steps: that the aqueous solution controlling 6-amino-penicillanic acid controls at 10～25 DEG C, uses
Ammonia regulation solution ph, to the isoelectric point, IP of 6-APA, obtains the crystallization of 6-APA, filters, washs, is dried；Wherein, described
The isoelectric point, IP of 6-amino-penicillanic acid be 4.3.
2. the preparation method of 6-amino-penicillanic acid as claimed in claim 1, it is characterised in that the temperature of described cracking reaction
Degree is for 26-38 DEG C, and in course of reaction, the pH of fermentation liquid is 7.0-8.5, and the response time of cracking reaction is 1-3h.
3. the preparation method of 6-amino-penicillanic acid as claimed in claim 1, it is characterised in that described microfiltration, ultrafiltration and
Filter membrane used in nanofiltration is organic membrane, ceramic membrane or metal film.
4. the preparation method of the 6-amino-penicillanic acid as described in claim 1 or 3, it is characterised in that filter in described ultrafiltration
The molecular cut off of film is 3000--50000 dalton.
5. the preparation method of the 6-amino-penicillanic acid as described in claim 1 or 3, in described nanofiltration, filter membrane dams point
Son amount is 100～260 dalton.
6. the preparation method of 6-amino-penicillanic acid as claimed in claim 1, it is characterised in that described 6-amino penicillium sp
The mass percent of 6-amino-penicillanic acid in mixed solution that concentrates of alkanoic acid and phenylacetic acid is 5%-15%.
7. the preparation method of 6-amino-penicillanic acid as claimed in claim 1, it is characterised in that de-in described desolventing technology
Toner is activated carbon, aluminium sesquioxide or a combination thereof, and described resin is hypercrosslinked polymeric resin or macroporous adsorbent resin.
8. the preparation method of 6-amino-penicillanic acid as claimed in claim 7, described hypercrosslinked polymeric resin model be AH,
One in NDa-150, the model of described macroporous adsorbent resin is AmberliteXAD-4.
9. claim 1-3 or 6-8 arbitrary as described in the preparation method of 6-amino-penicillanic acid, it is characterised in that it also wraps
Including phenylacetic acid De contamination step, wherein said phenylacetic acid De contamination comprises the steps: to use interpolation ethanol or the hydrogen of acetone
Sodium hydroxide solution washing is adsorbed with the resin of phenylacetic acid.
10. the preparation method of 6-amino-penicillanic acid as claimed in claim 4, it is characterised in that it also includes phenylacetic acid
De contamination step, wherein said phenylacetic acid De contamination comprises the steps: to use the sodium hydroxide adding ethanol or acetone molten
Liquid washing is adsorbed with the resin of phenylacetic acid.
The preparation method of 11. 6-amino-penicillanic acids as claimed in claim 5, it is characterised in that it also includes phenylacetic acid
De contamination step, wherein said phenylacetic acid De contamination comprises the steps: to use the sodium hydroxide adding ethanol or acetone molten
Liquid washing is adsorbed with the resin of phenylacetic acid.
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| CN105567778B (en) * | 2016-03-02 | 2019-03-12 | 青岛市市立医院 | A kind of preparation method of 6-amino-penicillanic acid |
| CN105693747B (en) * | 2016-03-07 | 2018-02-06 | 内蒙古常盛制药有限公司 | A kind of 6 APA is without the dry technology of solvent washing |
| CN105732663B (en) * | 2016-03-28 | 2018-03-16 | 西藏百年汉克药业有限公司 | The preparation method of 6 aminopenicillanic acids |
| CN109535179B (en) * | 2017-09-22 | 2020-05-22 | 联邦制药(内蒙古)有限公司 | Improved 6-APA extraction method |
| CN108658757B (en) * | 2018-05-11 | 2020-11-06 | 国药集团威奇达药业有限公司 | Method for recovering phenylacetic acid from 6-aminopenicillanic acid enzymatic aqueous solution |
| CN109970226B (en) * | 2019-04-16 | 2020-04-03 | 同舟纵横（厦门）流体技术有限公司 | Method for treating sodium phenylacetate deesterification liquid generated by cracking penicillin G sylvite crystallization mother liquor |
| CN113861221A (en) * | 2020-06-30 | 2021-12-31 | 伊犁川宁生物技术有限公司 | 6-aminopenicillanic acid production method |
| CN112047961B (en) * | 2020-08-13 | 2021-12-17 | 国药集团威奇达药业有限公司 | Method for separating and crystallizing 6-aminopenicillanic acid from penicillin enzymolysis liquid |
| CN113005168B (en) * | 2021-03-10 | 2022-05-17 | 济宁市技师学院 | Preparation method of 6-aminopenicillanic acid |
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| CN1314690C (en) * | 2005-06-08 | 2007-05-09 | 中国科学院过程工程研究所 | Coupling process for preparing intermediate of semi-synthetic Cephalosperin antibiotics from penicillin fermentation liquid |
| CN100543026C (en) * | 2005-10-14 | 2009-09-23 | 中国科学院过程工程研究所 | Ion liquid extraction penicillin and enzymic catalytic reaction coupling prepare the method for semisynthetic antibiotic 6-amino penicillanic acid |
| CN101735243A (en) * | 2008-11-11 | 2010-06-16 | 华北制药股份有限公司 | Process for preparing straight-through 6-aminopenicillanic acid |
| CN102925526B (en) * | 2012-11-23 | 2014-10-15 | 华北制药河北华民药业有限责任公司 | Preparation method for 6-amino penicillanic acid |
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