CN113214103A - Subsequent treatment method for enzymatic synthesis of D-p-hydroxyphenylglycine - Google Patents
Subsequent treatment method for enzymatic synthesis of D-p-hydroxyphenylglycine Download PDFInfo
- Publication number
- CN113214103A CN113214103A CN202110441874.0A CN202110441874A CN113214103A CN 113214103 A CN113214103 A CN 113214103A CN 202110441874 A CN202110441874 A CN 202110441874A CN 113214103 A CN113214103 A CN 113214103A
- Authority
- CN
- China
- Prior art keywords
- hydroxyphenylglycine
- feed liquid
- subsequent treatment
- oxidation
- enzymatic synthesis
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/40—Separation; Purification
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/07—Optical isomers
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by an enzymatic method, which relates to the field of chemical pharmacy and comprises the following steps: (1) and (3) oxidation: adjusting the pH value of a feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by an enzymatic method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid; (2) diluting: diluting the oxidized feed liquid with water to obtain D-p-hydroxyphenylglycine diluent; (3) and (3) decoloring: adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon to adsorb and remove impurities, and filtering the activated carbon to obtain a D-p-hydroxyphenylglycine decolored solution; (4) preparing a finished product: and D-p-hydroxyphenylglycine decolored solution is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and a finished product of D-p-hydroxyphenylglycine is obtained after filtration, washing and drying. The method has the advantages of simple process, good impurity removal effect and excellent finished product quality, and is suitable for industrial production.
Description
Technical Field
The invention relates to the technical field of chemical pharmacy, in particular to a subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by an enzymatic method.
Background
D-p-hydroxyphenylglycine (D-p-hydroxyphenylglycine, D-p-HPG, D-HPG) is an important intermediate for preparing semi-synthetic penicillin and semi-synthetic cephalosporin, and can be used for synthesizing broad-spectrum antibiotics of amoxicillin, cefadroxil, cefoperazone, cefaclor and the like.
The existing method for preparing D-p-hydroxyphenylglycine mainly comprises a biological enzyme catalysis method and a chemical synthesis method, wherein the biological enzyme catalysis method has the advantages of small environmental pollution and mild reaction conditions, and is gradually a research hotspot.
The preparation of D-p-hydroxyphenylglycine by a biological enzyme catalysis method mainly uses p-hydroxyphenylhydantoin (DL-HPH) as a substrate, and is converted into N-carbamoyl D-p-hydroxyphenylglycine (D-CH PG) by using D-hydantoinase (IDH), and then converted into D-p-hydroxyphenylglycine by using D-carbamoyl hydrolase (IDC). The enzyme conversion process is completed in an aqueous solution system, and the solid-liquid mixed feed liquid after the enzyme reaction contains soluble protein, organic pigment and other inorganic salt impurities besides the target product D-p-hydroxyphenylglycine, so that the target product needs to be extracted from the reaction system by adopting a proper method.
At present, few reports are reported on post-treatment processes for synthesizing D-p-hydroxyphenylglycine by enzymatic reaction, and the existing post-treatment methods comprise:
resin adsorption extraction method: adsorbing D-p-hydroxyphenylglycine by using resin, resolving to obtain a resolving solution, carrying out ultrafiltration decoloration and nanofiltration concentration on the resolving solution under an alkaline condition, and carrying out isoelectric point crystallization on the concentrated solution to obtain a finished product D-p-hydroxyphenylglycine. The process has the advantages of large resin amount, high cost, incomplete impurity removal and soluble protein residue, is only suitable for recovering a small amount of crystallization mother liquor, and causes unqualified alkaline absorbance and poor product quality.
The multi-stage filter membrane purification method comprises the following steps: firstly, recrystallizing and refining a solid crude product generated by an enzyme reaction, and purifying a crystallization mother liquor and a liquid crude product obtained after the enzyme reaction through multi-stage filter membrane filtration; in the purification process, soluble protein and organic pigment impurities are removed through ultrafiltration and nanofiltration, desalting is performed through an electric point of electrodialysis, and finally concentration, filtration, washing and drying are performed to obtain a finished product of D-p-hydroxyphenylglycine. The impurity removal of the filter membrane is difficult to be thorough, and the impurity residue is denatured and developed under the strong alkaline condition, so that the alkali absorbance is unqualified, and the product quality is influenced.
Therefore, how to provide a post-treatment method with simple process, good impurity removal effect and excellent finished product quality for the feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by an enzyme method becomes a technical problem to be solved in the field.
Disclosure of Invention
In view of the above, the invention provides a subsequent treatment method for enzymatic synthesis of D-p-hydroxyphenylglycine, which has the advantages of simple process, good impurity removal effect and excellent finished product quality, and is suitable for industrial production.
In order to achieve the purpose, the invention adopts the following technical scheme:
a subsequent treatment method for enzymatic synthesis of D-p-hydroxyphenylglycine comprises the following steps:
(1) oxidation by oxygen
Adjusting the pH value of a feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by an enzymatic method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid;
(2) dilution of
Diluting the oxidized feed liquid with water to obtain D-p-hydroxyphenylglycine diluent;
(3) decolorization of
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon to adsorb and remove impurities, and filtering the activated carbon to obtain a D-p-hydroxyphenylglycine decolored solution;
(4) to obtain the final product
And D-p-hydroxyphenylglycine decolored solution is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and a finished product of D-p-hydroxyphenylglycine is obtained after filtration, washing and drying.
Before the active carbon is used for decoloring, the impurities are oxidized under the alkaline condition to enable the non-developed soluble protein impurities to be developed and denatured, and then the impurities are diluted and adjusted to be acidic, so that the adsorption efficiency of the active carbon can be ensured, and the product with qualified alkali absorbance can be obtained. After decolorization, the D-p-hydroxyphenylglycine finished product with the purity of more than 99 percent can be obtained by reverse osmosis concentration, filtration, washing and drying, the process is simple, the environmental protection treatment difficulty is low, and the method is suitable for popularization and application.
Preferably, in the step (1),
adjusting pH to 10.0-10.2;
the alkali used for adjusting the pH is the same as the alkali used for the reaction for synthesizing the D-p-hydroxyphenylglycine by the enzyme method.
Preferably, in the step (1),
adding hydrogen peroxide until the color of the feed liquid is not changed.
The discharging liquid is colorless under the condition of normal discharging without contacting with air, if the discharging liquid is oxidized by air, the discharging liquid is light yellow, the discharging liquid is adjusted to be alkaline and is generally yellow or light brown, and the color is gradually deepened after the discharging liquid is oxidized by adding hydrogen peroxide and finally becomes brownish black. If the oxidation is not thorough, the active carbon cannot thoroughly decolorize the feed liquid to be colorless and transparent, and the decolorized liquid is light yellow, namely, impurities are not thoroughly removed, and the absorbance of the finished product alkali is influenced.
Preferably, in the step (2),
the oxidation feed liquid is diluted by 3 to 6 times.
Further preferably, the oxidation feed liquid is diluted 4 to 5 times.
Preferably, in the step (3),
the pH is adjusted to 5.0-5.2, and the weak acidity is more beneficial to subsequent decolorization and impurity removal;
the acid used for adjusting the pH is hydrochloric acid with the mass fraction of 20-30%.
Preferably, in the step (3),
the active carbon dosage is 1-5kg/m3D-p-hydroxyphenylglycine diluent is prepared from D-p-hydroxyphenylglycine,
the adsorption time is 20-60 min;
the slow stirring is carried out in the adsorption process, so as to ensure that the active carbon is not settled at the bottom of the tank, thereby achieving good adsorption effect.
Further preferably, the stirring speed is 60-80 r/min.
Preferably, in the step (4),
before reverse osmosis concentration, adding a reducing agent into the D-p-hydroxyphenylglycine decolored solution to neutralize redundant hydrogen peroxide;
the concentration multiple of reverse osmosis concentration is 10-20 times.
Preferably, the drying temperature in the step (4) is below 60 ℃.
Further preferably, the drying temperature in the step (4) is 50-55 ℃.
Preferably, the method further comprises:
(5) monovalent salt recovery
Carrying out nanofiltration on the filtrate obtained by filtering in the step (4) to obtain monovalent salt-containing dialysate and monovalent salt-removing feed liquid;
and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
Preferably, the method further comprises:
(6) trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid;
the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
Preferably, in the step (4),
returning RO dialysate obtained by reverse osmosis concentration to the step (2) for diluting the oxidation feed liquid or for nanofiltration; washing water obtained in the washing step is returned to the step (2) for diluting the oxidation feed liquid;
in the steps (5) and (6),
condensed water evaporated by concentration and evaporation is returned to the step (2) for diluting the oxidation feed liquid;
in the step (6), the step (c),
and (4) returning the material liquid except the trivalent salt to the step (3) for diluting the oxidation material liquid.
The monovalent salt and the trivalent salt are classified and recycled, and water generated in each process is reasonably used, so that the effective utilization of resources is realized.
Further preferably, in the step (5),
the pressure of the nanofiltration membrane is less than or equal to 2.5MPa, and the molecular weight cut-off of the nanofiltration membrane is 150-.
Further preferably, after desalting, the monovalent salt concentration is reduced by 5 to 10 times, and the trivalent salt concentration is reduced by 5 to 10 times.
In summary, the recrystallization process for removing the solid crude product in the invention intensively treats the reaction feed liquid, and reduces the process steps; in the process, salts generated by reaction and post-treatment are classified and recovered, and waste liquid generated in each process is reasonably reused, so that the environmental protection treatment difficulty is reduced, and reasonable utilization of resources is realized; the waste liquid after desalting is reused, so that the yield is improved to over 94 percent, and the production cost is reduced; the D-p-hydroxyphenylglycine obtained by the treatment of the process disclosed by the invention is bright white in appearance, the purity is more than 99%, and the indexes of finished products such as detection ratio, acid absorbance, alkali absorbance and the like are excellent.
Drawings
FIG. 1 shows a process flow diagram of the present invention.
Detailed Description
The technical solutions in the embodiments of the present invention are clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The liquid obtained after the enzymatic synthesis of D-p-hydroxyphenylglycine used in the examples is 2 batches, p-hydroxyphenylhydantoin is used as a raw material, and the liquid is prepared by converting D-hydantoinase and D-carbamoylhydrolase, wherein ammonia water and phosphoric acid are used for controlling the reaction process in the reaction process.
First batch enzymatic conversion 99.5%, 2m each3The volume of the feed liquid contains about 173.10kg of D-p-hydroxyphenylglycine.
The second batch had an enzymatic conversion of 99.6% per 2m3The volume of the feed liquid contains about 173.274kg of D-p-hydroxyphenylglycine.
Example 1
As shown in figure 1, the first batch of reacted feed liquid 2m after enzymatic synthesis of D-p-hydroxyphenylglycine3The treatment is carried out, and comprises the following steps:
(1) oxidation by oxygen
The pH value of the feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by the enzyme method is adjusted to 10.1 by using concentrated ammonia water, 10kg of hydrogen peroxide (the dosage can ensure that the oxidation feed liquid achieves the maximum oxidation effect through experiments) is added under the condition of slow stirring, and the oxidation feed liquid is oxidized for 2 hours to obtain the oxidation feed liquid.
(2) Dilution of
Transferring the oxidized material liquid into a dilution tank, adding water to dilute the oxidized material liquid to about 9.0m3And D-p-hydroxyphenylglycine diluent is obtained.
(3) Decolorization of
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to 5.0 by using hydrochloric acid with the mass fraction of 30%, adding 30kg of active carbon, slowly stirring (70r/min), adsorbing for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decoloration solution.
(4) To obtain the final product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorized solution to neutralize redundant hydrogen peroxide, and then concentrating the D-p-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 8.5m3And (3) the solution enters an RO dialysate storage tank (the solution can be returned to the step (2) to be used for diluting the next batch of oxidation feed liquid or used for subsequent nanofiltration); filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
d-p-hydroxyphenylglycine with the proportion of about 8 percent is contained in the filtrate, and the subsequent desalting treatment is carried out;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water contains about 4 percent of D-p-hydroxyphenylglycine and is used for the application of the next production dilution step; the wet powder is dried at 50 ℃ to obtain 150.94 kgD-p-hydroxyphenylglycine with the yield of 87.20 percent.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in the nanofiltration process is 1.24 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 2.73%.
Example 2
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3The treatment is carried out, and comprises the following steps:
(1) oxidation by oxygen
Adjusting the pH value of the feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by the enzyme method to 10.1 by using concentrated ammonia water, adding 10kg of hydrogen peroxide under the condition of slow stirring, and oxidizing for 2 hours to obtain oxidized feed liquid.
(2) Dilution of
The oxidation feed liquid was transferred to a dilution tank, diluted with the washing water, the condensed water and the feed liquid for removing trivalent salt obtained in example 1, and then diluted with water to about 9.5m3And D-p-hydroxyphenylglycine diluent is obtained.
(3) Decolorization of
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to 5.0 by using hydrochloric acid with the mass fraction of 30%, adding 30kg of active carbon, slowly stirring (70r/min), adsorbing for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decoloration solution.
(4) To obtain the final product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorized solution to neutralize redundant hydrogen peroxide, and then concentrating the D-p-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 9.0m3Entering an RO dialysate storage tank; filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water is used for indiscriminately applying the dilution step in the next batch of production; the wet powder is dried at 50 ℃ to obtain 164.40 kgD-p-hydroxyphenylglycine with yield of 94.88%.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in the nanofiltration process is 1.14 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 3.11%.
Example 3
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3The treatment is carried out, and comprises the following steps:
(1) oxidation by oxygen
Adjusting the pH of the feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by the enzyme method to 10.2 by using concentrated ammonia water, adding 10kg of hydrogen peroxide under the condition of slow stirring, and oxidizing for 2 hours to obtain oxidized feed liquid.
(2) Dilution of
The oxidation feed liquid was transferred to a dilution tank, diluted with the washing water, the condensed water and the feed liquid for removing trivalent salt obtained in example 2, and then diluted with water to about 9.5m3And D-p-hydroxyphenylglycine diluent is obtained.
(3) Decolorization of
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to 5.0 by using hydrochloric acid with the mass fraction of 30%, adding 30kg of active carbon, slowly stirring (70r/min), adsorbing for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decoloration solution.
(4) To obtain the final product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorized solution to neutralize redundant hydrogen peroxide, and then concentrating the D-p-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 9.0m3Entering an RO dialysate storage tank; filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water is used for indiscriminately applying the dilution step in the next batch of production; the wet powder is dried at 50 ℃ to obtain 164.974 kgD-p-hydroxyphenylglycine with yield of 95.21%.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The D-p-hydroxyphenylglycine loss in the nanofiltration process is 0.92 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 2.96%.
Comparative example 1
Taking the first batch of reacted feed liquid 2m after enzymatic synthesis of D-p-hydroxyphenylglycine3Performing multi-stage filter membrane impurity removal treatment as follows:
(1) refining
Filtering the feed liquid to perform solid-liquid separation, and adding 152.88kg of wet powder of 1.0m3Adding concentrated ammonia water into purified water to adjust pH to about 10.1, dissolving, adding active carbon 30kg, stirring for decolorizing for more than 30min, and filtering to remove active carbon. Adding 30% HCl into the decolorized feed liquid to adjust the pH to about 5.0, acidifying and crystallizing, stirring for crystal growth for 1h, and performing suction filtration to obtain a crystallization mother liquid and crystals; washing the crystal with about 60L of water twice, and vacuum drying at 50 ℃ to obtain 67.01kg of finished product with the yield of 38.71%; the washing water is used for the next batch production.
(2) Ultrafiltration
Removing impurities from the liquid material and crystallized mother liquor by ultrafiltration membrane, and concentrating with ultrafiltration membrane of 0.8m3Washing with water; the loss of D-p-hydroxyphenylglycine in the ultrafiltration process is 2.29%.
(3) Nanofiltration
Subjecting the ultrafiltered dialysate to ultrafiltration of about 3.8m3Decolorizing with nanofiltration membrane, and collecting the concentrated solution with a volume of 0.8m3Washing with water; the D-p-hydroxyphenylglycine loss in the nanofiltration process is 2.25 percent.
(4) RO concentration
Concentrating the nanofiltration decolorized solution through an RO membrane to obtain RO dialysate and RO concentrated solution;
the RO dialysate is discharged at about 4.0m3Entering an RO dialysate storage tank;
filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
d-p-hydroxyphenylglycine accounts for about 4.8% of the filtrate, and desalting treatment is carried out subsequently;
the crude product is soaked and washed twice with 100L of water, the proportion of D-p-hydroxyphenylglycine in the washing water is about 2.1 percent, and the D-p-hydroxyphenylglycine is used for the next secondary production; the crude product is washed by water and dried to obtain 85.42 kgD-p-hydroxyphenylglycine with the yield of 49.35 percent and the total yield of the finished product D-p-hydroxyphenylglycine of the batch of 88.06 percent.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The D-p-hydroxyphenylglycine loss in the nanofiltration process is 0.86 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt. The trivalent salt removing material liquid is used for the next batch of production.
Electrodialysis step D-p-hydroxyphenylglycine loss 2.47%.
Comparative example 2
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3Performing multi-stage filter membrane impurity removal treatment as follows:
(1) refining
Filtering the feed liquid to perform solid-liquid separation, and adding 158.41kg of wet powder of 1.0m3Adding concentrated ammonia water into purified water to adjust pH to about 10.1, dissolving, adding active carbon 30kg, stirring for decolorizing for more than 30min, and filtering to remove active carbon. Adding 30% HCl into the decolorized feed liquid to adjust the pH to about 5.0, acidifying and crystallizing, stirring for crystal growth for 1h, and performing suction filtration to obtain a crystallization mother liquid and crystals; washing the crystal with about 60L of water twice, and vacuum drying at 50 ℃ to obtain 66.53kg of finished product with the yield of 38.39%; the washing water is used for the next batch production.
(2) Ultrafiltration
Removing impurities from the liquid material and crystallized mother liquor by ultrafiltration membrane, and concentrating with ultrafiltration membrane of 0.8m3Washing with water; the loss of D-p-hydroxyphenylglycine in the ultrafiltration process was 2.37%.
(3) Nanofiltration
Subjecting the ultrafiltered dialysate to ultrafiltration of about 3.8m3And the washing water and the trivalent salt removing feed liquid in the comparative example 1 are about 4.6m in total3Decolorizing with nanofiltration membrane, and collecting the concentrated solution with a volume of 0.8m3Washing with water; the D-p-hydroxyphenylglycine loss in the nanofiltration process is 2.74 percent.
(4) RO concentration
Concentrating the nanofiltration decolorized solution through an RO membrane to obtain RO dialysate and RO concentrated solution;
the RO dialysate is discharged by about 5.1m3Entering an RO dialysate storage tank;
filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
the crude product is soaked and washed twice with 100L of water, the proportion of D-p-hydroxyphenylglycine in the washing water is about 2.1 percent, and the D-p-hydroxyphenylglycine is used for the next secondary production; the crude product is washed by water and dried to obtain 90.81 kgD-p-hydroxyphenylglycine with the yield of 52.41 percent and the total yield of the finished product D-p-hydroxyphenylglycine of the batch of 90.80 percent.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The D-p-hydroxyphenylglycine loss in the nanofiltration process is 0.91 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt. The trivalent salt removing material liquid is used for the next batch of production.
Electrodialysis step D-p-hydroxyphenylglycine loss was 2.30%.
Comparative example 3
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3The following treatment (no oxidation step) was carried out:
(1) decolorization pretreatment
Adjusting pH of the material liquid obtained by enzymatic synthesis of D-p-hydroxyphenylglycine to 10.1 with concentrated ammonia water, mechanically diluting condensate water, crude product washing water and trivalent salt removing material liquid in the previous qualified batches, and adding waterDiluting to about 9.5m3And D-p-hydroxyphenylglycine diluent is obtained.
(2) Decolorization of
And (3) adjusting the pH value of the D-p-hydroxyphenylglycine diluent to 5.0 by using hydrochloric acid with the mass fraction of 30%, adding 30kg of active carbon, stirring and adsorbing for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decoloration solution.
(3) To obtain the final product
Concentrating the D-p-hydroxyphenylglycine decolorized solution by an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 9.0m3Entering an RO dialysate storage tank; filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water is used for indiscriminately applying the dilution step in the next batch of production; the wet powder is dried at 50 ℃ to obtain 163.89 kgD-p-hydroxyphenylglycine with yield of 94.58%.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in the nanofiltration process is 1.17 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 3.03%.
Comparative example 4
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3The following treatments (decolorization by adding activated carbon under alkaline conditions) are carried out:
(1) oxidation by oxygen
Adjusting the pH value of the feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by the enzyme method to 10.1 by using concentrated ammonia water, adding 10kg of hydrogen peroxide under the condition of slow stirring, and oxidizing for 2 hours to obtain oxidized feed liquid.
(2) Dilution of
Transferring the oxidized material liquid into a dilution tank, diluting with washing water, condensed water and trivalent salt removing material liquid obtained from the previous qualified batches, and adding water to dilute to about 9.5m3And D-p-hydroxyphenylglycine diluent is obtained.
(3) Decolorization of
Adding 30kg of active carbon into the D-p-hydroxyphenylglycine diluent, stirring and adsorbing for 30min, and filtering the active carbon to obtain a D-p-hydroxyphenylglycine decolored solution.
(4) To obtain the final product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolored solution to neutralize redundant hydrogen peroxide, and then adjusting the pH of the D-p-hydroxyphenylglycine decolored solution to 5.0 by using hydrochloric acid with the mass fraction of 30%;
concentrating the D-p-hydroxyphenylglycine decolorized solution by an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 9.0m3Entering an RO dialysate storage tank; filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water is used for indiscriminately applying the dilution step in the next batch of production; the wet powder is dried at 50 ℃ to obtain 164.454 kgD-p-hydroxyphenylglycine with yield of 94.91%.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in the nanofiltration process is 1.35 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 2.93%.
Comparative example 5
Taking the reacted feed liquid of 2m after the second batch of enzymatic synthesis of D-p-hydroxyphenylglycine3The following treatments were carried out (oxidative decolorization under weakly acidic conditions):
(1) decolorization of
Transferring the feed liquid into a dilution tank, adjusting pH to 5.0 with 30% hydrochloric acid, diluting the feed liquid with washing water, condensate water and trivalent salt-removing feed liquid obtained from the previous qualified batches, and adding water to dilute the feed liquid to about 9.5m3Completely dissolving the materials and keeping the pH value at 5.0; adding 10kg of hydrogen peroxide under the condition of slow stirring, and oxidizing for 2 hours; then adding 30kg of active carbon, stirring and adsorbing for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decolored solution.
(2) To obtain the final product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorized solution to neutralize redundant hydrogen peroxide, and then concentrating the D-p-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and an RO concentrated solution; the RO dialysate is discharged by about 9.0m3Entering an RO dialysate storage tank; filtering the RO concentrated solution to obtain a D-p-hydroxyphenylglycine crude product and a filtrate;
soaking and washing the crude product of D-p-hydroxyphenylglycine with purified water for 2 times, and using 160L of water each time; the washing water is used for indiscriminately applying the dilution step in the next batch of production; the wet powder is dried at 50 ℃ to obtain 164.75 kgD-p-hydroxyphenylglycine with the yield of 95.08%.
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) by using a nanofiltration membrane with the molecular weight cutoff of 150-; and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in the nanofiltration process is 1.33 percent.
(6) Trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid; the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
And (5) concentrating the condensed water evaporated by evaporation and the trivalent salt removal feed liquid in the steps (5) and (6) and returning the condensed water and the trivalent salt removal feed liquid to the step (2) for diluting the next batch of oxidation feed liquid.
Electrodialysis step D-p-hydroxyphenylglycine loss 3.02%.
D-p-hydroxyphenylglycine prepared in examples 1 to 3 and comparative examples 1 to 5 was examined, and the results are shown in Table 1.
TABLE 1
As can be seen from the data in the table 1, the impurities influencing the alkali absorbance are not completely removed in the comparative examples 1 and 2 by using a multistage filter membrane, and finally the alkali absorbance is higher and cannot reach the qualified standard; and the solid and liquid crude products are separately treated, so that the treatment process is increased, the loss is more, and the yield is still about 3 percent lower than that of the method after the feed liquid is reused.
Comparative examples 3-5 the oxidation, dilution and decoloration processes are slightly different from those of the examples, but the quality of the finished product is poor; it can be seen that the oxidation, dilution and decolorization steps must be processed in the process sequence of the present invention.
The general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
Claims (10)
1. A subsequent treatment method for enzymatic synthesis of D-p-hydroxyphenylglycine is characterized by comprising the following steps:
(1) oxidation by oxygen
Adjusting the pH value of a feed liquid obtained by the reaction of synthesizing D-p-hydroxyphenylglycine by an enzymatic method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid;
(2) dilution of
Diluting the oxidized feed liquid with water to obtain D-p-hydroxyphenylglycine diluent;
(3) decolorization of
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon to adsorb and remove impurities, and filtering the activated carbon to obtain a D-p-hydroxyphenylglycine decolored solution;
(4) to obtain the final product
And D-p-hydroxyphenylglycine decolored solution is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and a finished product of D-p-hydroxyphenylglycine is obtained after filtration, washing and drying.
2. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (1), the step (c),
adjusting pH to 10.0-10.2;
the alkali used for adjusting the pH is the same as the alkali used for the reaction for synthesizing the D-p-hydroxyphenylglycine by the enzyme method.
3. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (1), the step (c),
adding hydrogen peroxide until the color of the feed liquid is not changed.
4. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (2),
the oxidation feed liquid is diluted by 3 to 6 times.
5. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (3), the step (c),
adjusting pH to 5.0-5.2;
the acid used for adjusting the pH is hydrochloric acid with the mass fraction of 20-30%.
6. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (3), the step (c),
the active carbon dosage is 1-5kg/m3D-p-hydroxyphenylglycine diluent is prepared from D-p-hydroxyphenylglycine,
the adsorption time is 20-60 min.
7. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
in the step (4), the step (c),
before reverse osmosis concentration, adding a reducing agent into the D-p-hydroxyphenylglycine decolored solution to neutralize redundant hydrogen peroxide;
the concentration multiple of reverse osmosis concentration is 10-20 times.
8. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 1,
further comprising:
(5) monovalent salt recovery
Carrying out nanofiltration on the filtrate obtained by filtering in the step (4) to obtain monovalent salt-containing dialysate and monovalent salt-removing feed liquid;
and concentrating the dialyzate containing monovalent salt and evaporating to dryness to obtain monovalent salt.
9. The method of claim 8, wherein the subsequent treatment of D-p-hydroxyphenylglycine is obtained by enzymatic synthesis,
further comprising:
(6) trivalent salt recovery
Performing electrodialysis on the monovalent salt removing feed liquid obtained in the step (5) to obtain trivalent salt brine and trivalent salt removing feed liquid;
the trivalent salt brine is concentrated and evaporated to dryness to prepare trivalent salt.
10. The subsequent treatment method for the enzymatic synthesis of D-p-hydroxyphenylglycine according to claim 9,
in the step (4), the step (c),
returning RO dialysate obtained by reverse osmosis concentration to the step (2) for diluting the oxidation feed liquid or for nanofiltration; washing water obtained in the washing step is returned to the step (2) for diluting the oxidation feed liquid;
in the steps (5) and (6),
condensed water evaporated by concentration and evaporation is returned to the step (2) for diluting the oxidation feed liquid;
in the step (6), the step (c),
and (3) returning the material liquid except the trivalent salt to the step (2) for diluting the oxidation material liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110441874.0A CN113214103B (en) | 2021-04-23 | 2021-04-23 | Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110441874.0A CN113214103B (en) | 2021-04-23 | 2021-04-23 | Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113214103A true CN113214103A (en) | 2021-08-06 |
| CN113214103B CN113214103B (en) | 2023-05-02 |
Family
ID=77088948
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110441874.0A Active CN113214103B (en) | 2021-04-23 | 2021-04-23 | Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113214103B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116730856A (en) * | 2023-06-19 | 2023-09-12 | 浙江竹子制药有限公司 | Synthesis method of tranexamic acid |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884144A (en) * | 2006-06-29 | 2006-12-27 | 南京大学 | Method for treating waste water in production of para-hydroxyphenyl hydantoin |
| CN104357528A (en) * | 2014-10-29 | 2015-02-18 | 国药集团威奇达药业有限公司 | Method for comprehensively recovering effective ingredients in amoxicillin mother liquid prepared by enzyme process |
| CN105254520A (en) * | 2015-10-12 | 2016-01-20 | 国药集团威奇达药业有限公司 | Recovery method for D-hydroxyphenylglycine in amoxicillin crystallization mother liquor of enzymatic synthesis |
| CN105838771A (en) * | 2015-01-16 | 2016-08-10 | 上海凯鑫分离技术股份有限公司 | Recovery process of amoxicillin crystal mother solution |
| WO2016202252A1 (en) * | 2015-06-16 | 2016-12-22 | 山西新宝源制药有限公司 | Method for synthesizing d-para-hydroxyphenylglycine methyl ester |
| CN111393315A (en) * | 2020-02-20 | 2020-07-10 | 国药集团威奇达药业有限公司 | Method for recovering D-p-hydroxyphenylglycine in D-p-hydroxyphenylglycine crystallization mother liquor synthesized by enzyme method |
-
2021
- 2021-04-23 CN CN202110441874.0A patent/CN113214103B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1884144A (en) * | 2006-06-29 | 2006-12-27 | 南京大学 | Method for treating waste water in production of para-hydroxyphenyl hydantoin |
| CN104357528A (en) * | 2014-10-29 | 2015-02-18 | 国药集团威奇达药业有限公司 | Method for comprehensively recovering effective ingredients in amoxicillin mother liquid prepared by enzyme process |
| CN105838771A (en) * | 2015-01-16 | 2016-08-10 | 上海凯鑫分离技术股份有限公司 | Recovery process of amoxicillin crystal mother solution |
| WO2016202252A1 (en) * | 2015-06-16 | 2016-12-22 | 山西新宝源制药有限公司 | Method for synthesizing d-para-hydroxyphenylglycine methyl ester |
| CN105254520A (en) * | 2015-10-12 | 2016-01-20 | 国药集团威奇达药业有限公司 | Recovery method for D-hydroxyphenylglycine in amoxicillin crystallization mother liquor of enzymatic synthesis |
| CN111393315A (en) * | 2020-02-20 | 2020-07-10 | 国药集团威奇达药业有限公司 | Method for recovering D-p-hydroxyphenylglycine in D-p-hydroxyphenylglycine crystallization mother liquor synthesized by enzyme method |
Non-Patent Citations (1)
| Title |
|---|
| 戴连科等: "纳滤膜在D-对羟基苯甘氨酸提取中的应用", 《中国医药工业杂志》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116730856A (en) * | 2023-06-19 | 2023-09-12 | 浙江竹子制药有限公司 | Synthesis method of tranexamic acid |
| CN116730856B (en) * | 2023-06-19 | 2024-06-07 | 浙江竹子制药有限公司 | Synthesis method of tranexamic acid |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113214103B (en) | 2023-05-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| WO2011052824A1 (en) | Process for economically manufacturing xylose from hydrolysate using electrodialysis and direct recovery method | |
| CN108822163B (en) | Comprehensive cyclic production method of D-glucosamine hydrochloride | |
| CN102363594B (en) | Method for separating and purifying succinic acid from fermentation broth | |
| CN101486637A (en) | Method for extracting amber acid from fermentation liquor | |
| CN110272461B (en) | Method for purifying beta-thymidine from fermentation liquor | |
| CN106831894B (en) | A kind of method of deacetylation Coupling Adsorption separation D-Glucosamine Hydrochloride | |
| CN109081844B (en) | Method for extracting spectinomycin from fermentation culture | |
| CN111039808A (en) | Method for extracting tyrosine from fermentation liquor | |
| CN113214103B (en) | Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method | |
| CN109970543B (en) | Method for recycling acyclovir condensation by-product and solvent | |
| CN109369731B (en) | Method for removing glucose in xylose production process | |
| CN114436816B (en) | Method for efficiently extracting shikimic acid by ion exchange technology | |
| EP0778256A1 (en) | Process for producing an organic acid | |
| KR101073726B1 (en) | Economical manufacturing process of xylose from biomass hydrolysate using electrodialysis and direct recovery method | |
| CN104513151B (en) | A kind of method extracting high purity butylene diacid from succinate fermentation liquid | |
| CN109534983B (en) | Improved method for recovering phenylacetic acid waste liquid in 6-APA production | |
| CN113135965A (en) | System and method for producing crystalline xylose by using xylose mother liquor | |
| CN1207212C (en) | Production of potassium containing chemical products from waste liquid of molasses alcohol production | |
| CN111646638A (en) | Method for deep treatment and desalination of sucralose wastewater | |
| CN115583892B (en) | Preparation method of high-quality beta-alanine and impurity removal method | |
| CN112978993B (en) | Method for recovering thiocyanate ions from rectification waste liquid of erythromycin thiocyanate crystallization mother liquor and application | |
| CN113354702B (en) | Separation and purification process of high-purity NADP | |
| CN109234268B (en) | Method for treating yeast ribonucleic acid mother liquor | |
| CN109206313B (en) | Method for preparing succinic acid by succinate anaerobic fermentation liquid | |
| CN116574061A (en) | Method for separating and purifying L-histidine for cell culture medium |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |