CN113214103B - Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method - Google Patents
Subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by using enzymatic method Download PDFInfo
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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 of: (1) oxidation: adjusting the pH value of a feed liquid obtained by reacting the D-p-hydroxyphenylglycine synthesized by an enzyme method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid; (2) dilution: adding water into the oxidation feed liquid for dilution to obtain D-p-hydroxyphenylglycine diluent; (3) decoloring: adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon for adsorption and impurity removal, and filtering the activated carbon to obtain D-p-hydroxyphenylglycine decolorized solution; (4) preparing a finished product: the D-p-hydroxyphenylglycine decolorization liquid is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and the D-p-hydroxyphenylglycine decolorization liquid is filtered, washed and dried to obtain the finished product D-p-hydroxyphenylglycine. The invention has simple process, good impurity removing effect and excellent quality of finished products, 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 enzyme method.
Background
D-p-hydroxyphenylglycine (D-p-hydoxyphenylglycine, D-p-HPG, D-HPG) is an important intermediate for the production of semisynthetic penicillin and semisynthetic cephalosporins, and can be used for the synthesis of the broad-spectrum antibiotics amoxicillin, cefadroxil, cefoperazone, cefprozil, etc.
The current method for preparing the D-p-hydroxyphenylglycine mainly comprises a biological enzyme catalysis method and a chemical synthesis method, and the biological enzyme catalysis method has small environmental pollution and mild reaction conditions and gradually becomes a research hot spot.
The biological enzyme catalysis method for preparing the D-p-hydroxyphenylglycine mainly takes p-hydroxyphenylhydantoin (DL-HPH) as a substrate, utilizes D-hydantoin enzyme (IDH) to convert the D-hydroxyphenylglycine into N-carbamoyl D-p-hydroxyphenylglycine (D-CH PG), and utilizes D-carbamoyl hydrolase (IDC) to convert the N-carbamoyl D-p-hydroxyphenylglycine into the D-p-hydroxyphenylglycine. The enzyme conversion process is completed in an aqueous solution system, and the solid-liquid mixed solution 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.
The current post-treatment process for synthesizing D-p-hydroxyphenylglycine by enzymatic reaction has few reports, and the existing post-treatment method comprises the following steps:
resin adsorption extraction method: adsorbing the D-p-hydroxyphenylglycine by using resin, analyzing to obtain an analysis solution, carrying out ultrafiltration decolorization and nanofiltration concentration on the analysis solution under an alkaline condition, and carrying out isoelectric point crystallization on the concentrated solution to obtain the finished product D-p-hydroxyphenylglycine. The process has the advantages of large resin amount, high cost, incomplete impurity removal, and residual soluble protein, and is only suitable for recycling a small amount of crystallization mother liquor, so that the alkaline absorbance is unqualified, and the product quality is poor.
Multi-stage filter membrane purification method: firstly, recrystallizing and refining a solid crude product generated by the enzyme reaction, and purifying the crystallization mother liquor and the liquid crude product after the enzyme reaction together by multi-stage filter membrane filtration; in the purification process, soluble protein and organic pigment impurities are removed through ultrafiltration and nanofiltration, then the isoelectric point of electrodialysis is used for desalting, finally concentration is carried out, and the finished product D-p-hydroxyphenylglycine is obtained through filtration, washing and drying. The filter membrane is difficult to thoroughly remove impurities, and the impurities remain in a strong alkaline condition to be denatured and developed, 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 product quality aiming at the feed liquid obtained by the reaction of synthesizing the D-p-hydroxyphenylglycine by an enzyme method is 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 synthesizing D-p-hydroxyphenylglycine by an enzymatic method, which has the advantages of simple process, good impurity removal effect and excellent product quality, and is suitable for industrial production.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
a subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by an enzymatic method comprises the following steps:
(1) Oxidation
Adjusting the pH value of a feed liquid obtained by reacting the D-p-hydroxyphenylglycine synthesized by an enzyme method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid;
(2) Dilution of
Adding water into the oxidation feed liquid for dilution to obtain D-p-hydroxyphenylglycine diluent;
(3) Decoloring (decoloring)
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon for adsorption and impurity removal, and filtering the activated carbon to obtain D-p-hydroxyphenylglycine decolorized solution;
(4) Obtaining the finished product
The D-p-hydroxyphenylglycine decolorization liquid is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and the D-p-hydroxyphenylglycine decolorization liquid is filtered, washed and dried to obtain the finished product D-p-hydroxyphenylglycine.
Before the active carbon is used for decoloring, impurities are firstly oxidized under alkaline conditions to cause non-chromogenic soluble protein impurities to be chromogenic and denatured, and then diluted and adjusted to be acidic, so that the adsorption efficiency of the active carbon can be ensured, and a product with qualified alkali absorbance can be obtained. The D-p-hydroxyphenylglycine finished product with the purity of more than 99 percent can be obtained through reverse osmosis concentration, filtration, washing and drying after decolorization, 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),
the pH is adjusted to 10.0-10.2;
the alkali used for adjusting the pH is the same as the alkali used for the reaction of synthesizing the D-p-hydroxyphenylglycine by an enzyme method.
Preferably, in the step (1),
hydrogen peroxide is added until the feed liquid is not discolored any more.
The material liquid is colorless under the condition of no contact with air, and appears to be light yellow if oxidized by air, the material liquid is generally yellow or light brown after being regulated to be alkaline, and the color is gradually deepened after being oxidized by adding hydrogen peroxide, and finally becomes brownish black. If the oxidization is incomplete, the activated carbon cannot completely decolorize the feed liquid to be colorless and transparent, and the decolorized liquid has light yellow color, namely, the impurity is not completely removed, so that the absorbance of finished alkali can be influenced.
Preferably, in the step (2),
diluting the oxidation feed liquid by 3-6 times.
Further preferably, the oxidation feed solution is diluted 4-5 times.
Preferably, in the step (3),
the pH is adjusted to 5.0-5.2, so that the subsequent decolorization and impurity removal are more facilitated by weak acidity;
the acid used for adjusting the pH is hydrochloric acid with the mass fraction of 20-30%.
Preferably, in the step (3),
the dosage of the activated carbon is 1-5kg/m 3 D-p-hydroxyphenylglycine diluent,
the adsorption time is 20-60min;
and the activated carbon is slowly stirred in the adsorption process, so that the activated carbon is ensured not to be settled at the bottom of the tank, and a good adsorption effect is achieved.
Further preferably, the stirring speed is 60-80r/min.
Preferably, in the step (4),
adding a reducing agent into the D-p-hydroxyphenylglycine decolorization liquid before reverse osmosis concentration to neutralize redundant hydrogen peroxide;
the reverse osmosis concentration multiple is 10-20 times.
Preferably, the drying temperature in the step (4) is below 60 ℃.
Further preferably, the drying temperature in step (4) is 50-55 ℃.
Preferably, the method further comprises:
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) to obtain a monovalent salt-containing dialysate and monovalent salt-removing liquid;
concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
Preferably, the method further comprises:
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid;
concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
Preferably, in the step (4),
the RO dialysate obtained by reverse osmosis concentration is returned to the step (2) for diluting the oxidation liquid or for nanofiltration; the 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),
condensing, evaporating and drying condensed water, and returning the condensed water to the step (2) for diluting the oxidation feed liquid;
in the step (6), the step of (c),
the trivalent salt removal liquid returns to the step (3) for diluting the oxidation liquid.
The monovalent salt and the trivalent salt are classified and recovered, and water generated in each procedure is reasonably used, so that the effective utilization of resources is realized.
Further preferably, in the step (5),
the nanofiltration membrane inlet pressure is less than or equal to 2.5MPa, and the nanofiltration membrane interception molecular weight is 150-200.
Further preferably, the monovalent salt concentration is reduced by a factor of 5 to 10 and the trivalent salt concentration is reduced by a factor of 5 to 10 after desalting.
In summary, the recrystallization process for removing the solid crude product in the invention is to intensively treat the reaction feed liquid, so that the process steps are eliminated; the salt generated by the reaction and the post-treatment is classified and recycled in the process, the waste liquid generated by each procedure is reasonably applied, the environmental-friendly treatment difficulty is reduced, and the reasonable utilization of resources is realized; the waste liquid after desalting is used mechanically, the yield is increased to more than 94%, and the production cost is reduced; the D-p-hydroxyphenylglycine obtained by the process provided by the invention has the advantages of bright white appearance, purity of over 99%, and excellent indexes such as detection ratio, acid absorbance, alkali absorbance and the like.
Drawings
Figure 1 shows a process flow diagram of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely, and it is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The material liquid obtained after the enzymatic synthesis of D-p-hydroxyphenylglycine in the example is 2 batches, p-hydroxyphenylglycine is used as a raw material, D-hydantoin enzyme and D-carbamoyl hydrolase are used for conversion treatment, and ammonia water and phosphoric acid are used for controlling the reaction process in the reaction process.
The conversion rate of the first batch of enzyme reaction is 99.5 percent, per 2m 3 The volume of the feed liquid contains about 173.10kg of D-p-hydroxyphenylglycine.
The conversion rate of the second enzyme reaction is 99.6 percent, per 2m 3 The volume of the feed liquid contains about 173.274kg of D-p-hydroxyphenylglycine.
Example 1
As shown in FIG. 1, the first batch of material liquid 2m after the reaction of synthesizing D-p-hydroxyphenylglycine by the enzyme method 3 The processing comprises the following steps:
(1) Oxidation
And (3) regulating the pH value of the feed liquid obtained by the enzymatic synthesis of the D-p-hydroxyphenylglycine to 10.1 by using concentrated ammonia water, adding 10kg of hydrogen peroxide under the condition of slow stirring (the dosage can ensure that the oxidation feed liquid reaches the maximum oxidation effect through experiments), and oxidizing for 2 hours to obtain the oxidation feed liquid.
(2) Dilution of
Transferring the oxidation feed liquid into a dilution tank, adding water to dilute to about 9.0m 3 D-p-hydroxyphenylglycine diluent is obtained.
(3) Decoloring (decoloring)
Adjusting the pH 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 (70 r/min) for adsorption for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decolorized solution.
(4) Obtaining the finished product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorization liquid to neutralize redundant hydrogen peroxide, and concentrating the D-p-hydroxyphenylglycine decolorization liquid through an RO membrane to obtain RO dialysate and RO concentrated liquid; the RO dialysate was discharged at about 8.5m 3 Feeding into an RO dialysate tank (which may be returned to step (2) for dilution of the next batch of oxygenated feed solution, or 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% in the filtrate is subjected to subsequent desalting treatment;
soaking and washing the crude D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; the washing water has the proportion of about 4 percent of D-p-hydroxyphenylglycine and is used for the application of the next batch of production dilution steps; 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 1.24%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost by 2.73%.
Example 2
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The processing comprises the following steps:
(1) Oxidation
And (3) regulating the pH value of a feed liquid obtained by the enzymatic synthesis of D-p-hydroxyphenylglycine 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 an oxidized feed liquid.
(2) Dilution of
The oxidation feed solution was transferred to a dilution tank, diluted with the washing water, condensed water and trivalent salt-removed feed solution obtained in example 1, and then diluted to about 9.5m with water 3 D-p-hydroxyphenylglycine diluent is obtained.
(3) Decoloring (decoloring)
Adjusting the pH 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 (70 r/min) for adsorption for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decolorized solution.
(4) Obtaining the finished product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorization liquid to neutralize redundant hydrogen peroxide, and concentrating the D-p-hydroxyphenylglycine decolorization liquid through an RO membrane to obtain RO dialysate and RO concentrated liquid; the RO dialysate was discharged at about 9.0m 3 Feeding into 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 D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; washing water is used for carrying out the application of the next batch of production dilution step; the wet powder is dried at 50 ℃ to obtain 164.40 kgD-p-hydroxyphenylglycine with the yield of 94.88 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 1.14%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost 3.11%.
Example 3
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The processing comprises the following steps:
(1) Oxidation
And (3) regulating the pH value of a feed liquid obtained by the enzymatic synthesis of D-p-hydroxyphenylglycine 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 an oxidized feed liquid.
(2) Dilution of
The oxidation feed solution was transferred to a dilution tank, diluted with the washing water, condensed water and trivalent salt-removed feed solution obtained in example 2, and then diluted to about 9.5m with water 3 D-p-hydroxyphenylglycine diluent is obtained.
(3) Decoloring (decoloring)
Adjusting the pH 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 (70 r/min) for adsorption for 30min, and filtering the active carbon to obtain the D-p-hydroxyphenylglycine decolorized solution.
(4) Obtaining the finished product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorization liquid to neutralize redundant hydrogen peroxide, and concentrating the D-p-hydroxyphenylglycine decolorization liquid through an RO membrane to obtain RO dialysate and RO concentrated liquid; the RO dialysate was discharged at about 9.0m 3 Feeding into 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 D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; washing water is used for carrying out the application of the next batch of production dilution step; the wet powder is dried at 50 ℃ to obtain 164.974 kgD-p-hydroxyphenylglycine with the yield of 95.21 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 0.92%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost by 2.96%.
Comparative example 1
Taking 2m of feed liquid after the first batch of enzymatic synthesis of D-p-hydroxyphenylglycine 3 The following multistage filter membrane impurity removal treatment is carried out:
(1) Refining
Filtering the feed liquid to separate solid from liquid, filtering to obtain 152.88kg wet powder, and adding into 1.0m 3 Adding concentrated ammonia water into purified water to adjust pH to about 10.1, dissolving, adding activated carbon 30kg, decolorizing for more than 30min under stirring, and filtering to remove activated carbon. Adding 30% HCl into the decolorized feed liquid, regulating the pH to about 5.0, acidifying and crystallizing, stirring and crystallizing for 1h, and suction filtering to obtain crystallization mother liquor and crystals; the crystal is washed twice by 60L of water and dried in vacuum at 50 ℃ to obtain 67.01kg of finished product with the yield of 38.71%; washing water used for next batch production sleeveIs used.
(2) Ultrafiltration
Removing impurities from the liquid material and crystallization mother liquor after solid-liquid separation by ultrafiltration membrane, concentrating the ultrafiltration solution by 0.8m 3 Washing with water; the loss of D-p-hydroxyphenylglycine during ultrafiltration was 2.29%.
(3) Nanofiltration
Ultrafiltering the dialysate by about 3.8m 3 Decolorizing with nanofiltration membrane, concentrating the nanofiltration solution with 0.8m 3 Washing with water; the loss of D-p-hydroxyphenylglycine in nanofiltration process is 2.25%.
(4) RO concentration
Concentrating the nanofiltration decolorized solution through an RO membrane to obtain RO dialysate and RO concentrated solution;
the RO dialysate was discharged at about 4.0m 3 Feeding into 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 the subsequent desalting treatment is carried out;
the crude product is soaked and washed twice with 100L of water each time, and the D-p-hydroxyphenylglycine in the washing water accounts for about 2.1 percent and is used for the next batch production and application; the crude product is washed with 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 products 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 0.86%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt. The trivalent salt removing liquid is used for the next batch production and application.
Electrodialysis step D-p-hydroxyphenylglycine was lost by 2.47%.
Comparative example 2
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The following multistage filter membrane impurity removal treatment is carried out:
(1) Refining
Filtering the feed liquid to separate solid from liquid, filtering to obtain 158.41kg wet powder, and adding into 1.0m 3 Adding concentrated ammonia water into purified water to adjust pH to about 10.1, dissolving, adding activated carbon 30kg, decolorizing for more than 30min under stirring, and filtering to remove activated carbon. Adding 30% HCl into the decolorized feed liquid, regulating the pH to about 5.0, acidifying and crystallizing, stirring and crystallizing for 1h, and suction filtering to obtain crystallization mother liquor and crystals; the crystal is washed twice by 60L of water and dried in vacuum 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 crystallization mother liquor after solid-liquid separation by ultrafiltration membrane, concentrating the ultrafiltration solution by 0.8m 3 Washing with water; the loss of D-p-hydroxyphenylglycine during ultrafiltration was 2.37%.
(3) Nanofiltration
Ultrafiltering the dialysate by about 3.8m 3 And the total of the washing water, trivalent salt removed liquid in comparative example 1 was about 4.6m 3 Decolorizing with nanofiltration membrane, concentrating the nanofiltration solution with 0.8m 3 Washing with water; the loss of D-p-hydroxyphenylglycine in nanofiltration process is 2.74%.
(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 5.1m 3 Feeding into 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 each time, and the D-p-hydroxyphenylglycine in the washing water accounts for about 2.1 percent and is used for the next batch production and application; the crude product is washed with 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 products is 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 0.91%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt. The trivalent salt removing liquid is used for the next batch production and application.
Electrodialysis step D-p-hydroxyphenylglycine was lost by 2.30%.
Comparative example 3
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The following treatments (no oxidation step) were performed:
(1) Pretreatment for decoloring
The pH value of the feed liquid obtained by the enzymatic synthesis of D-p-hydroxyphenylglycine is adjusted to 10.1 by using strong ammonia water, condensed water, crude washing water and trivalent salt removal feed liquid in the prior qualified batch are mechanically diluted in the batch, and then water is added for dilution to about 9.5m 3 D-p-hydroxyphenylglycine diluent is obtained.
(2) Decoloring (decoloring)
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 activated carbon, stirring and adsorbing for 30min, and filtering the activated carbon to obtain the D-p-hydroxyphenylglycine decolorized solution.
(3) Obtaining the finished product
Concentrating the D-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and RO concentrated solution; the RO dialysate was discharged at about 9.0m 3 Feeding into 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 D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; washing water is used for carrying out the application of the next batch of production dilution step; the wet powder is dried at 50 ℃ to obtain 163.89 kgD-p-hydroxyphenylglycine with the yield of 94.58 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 1.17%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost 3.03%.
Comparative example 4
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The following treatments (decolorizing with activated carbon under alkaline conditions) were carried out:
(1) Oxidation
And (3) regulating the pH value of a feed liquid obtained by the enzymatic synthesis of D-p-hydroxyphenylglycine 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 an oxidized feed liquid.
(2) Dilution of
Transferring the oxidized feed liquid into a dilution tank, diluting with washing water, condensed water and trivalent salt-removed feed liquid obtained in the past qualified batch, and then adding water to dilute to about 9.5m 3 D-p-hydroxyphenylglycine diluent is obtained.
(3) Decoloring (decoloring)
Adding 30kg of activated carbon into the D-p-hydroxyphenylglycine diluent, stirring and adsorbing for 30min, and filtering the activated carbon to obtain the D-p-hydroxyphenylglycine decolorized solution.
(4) Obtaining the finished product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorization liquid to neutralize excessive hydrogen peroxide, and then adjusting the pH value of the D-p-hydroxyphenylglycine decolorization liquid to 5.0 by using hydrochloric acid with the mass fraction of 30%;
concentrating the D-hydroxyphenylglycine decolorized solution through an RO membrane to obtain RO dialysate and RO concentrated solution; the RO dialysate was discharged at about 9.0m 3 Feeding into 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 D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; washing water is used for carrying out the application of the next batch of production dilution step; the wet powder is dried at 50 ℃ to obtain 164.454 kgD-p-hydroxyphenylglycine with the yield of 94.91 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 1.35%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost by 2.93%.
Comparative example 5
Taking 2m of feed liquid after the reaction of synthesizing D-p-hydroxyphenylglycine by a second batch of enzymatic method 3 The following treatments (oxidative decolorization under weakly acidic conditions) were carried out:
(1) Decoloring (decoloring)
Transferring the feed liquid into a dilution tank, and regulating pH by using hydrochloric acid with mass fraction of 30%5.0, diluting the feed liquid by using the washing water, condensed water and trivalent salt removal feed liquid obtained in the past qualified batch, and then adding water to dilute to about 9.5m 3 The material was completely dissolved and maintained at pH 5.0; under the condition of slow stirring, adding 10kg of hydrogen peroxide, 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 decolorized solution.
(2) Obtaining the finished product
Adding a small amount of reducing agent into the D-p-hydroxyphenylglycine decolorization liquid to neutralize redundant hydrogen peroxide, and concentrating the D-p-hydroxyphenylglycine decolorization liquid through an RO membrane to obtain RO dialysate and RO concentrated liquid; the RO dialysate was discharged at about 9.0m 3 Feeding into 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 D-p-hydroxyphenylglycine product with purified water for 2 times, wherein 160L of water is used each time; washing water is used for carrying out the application of the next batch of production dilution step; the wet powder is dried at 50 ℃ to obtain 164.75 kgD-p-hydroxyphenylglycine with the yield of 95.08 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 cut-off of 150-200, the pressure of nanofiltration membrane is less than or equal to 2.5MPa, and monovalent salt-containing dialysate is discharged while water (RO dialysate is preferentially used) is fed for dilution, so that monovalent salt-containing dialysate and monovalent salt-removing liquid are finally obtained; concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt.
The loss of D-p-hydroxyphenylglycine in nanofiltration process is 1.33%.
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt.
And (5) concentrating, evaporating and drying the distilled condensate water in the steps (5) and (6), and returning the trivalent salt removal liquid to the step (2) for diluting the next batch of oxidation liquid.
Electrodialysis step D-p-hydroxyphenylglycine was lost 3.02%.
The D-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 table 1, the comparative examples 1 and 2 use multistage filter membranes to remove impurities, and the impurities affecting the alkali absorbance are not thoroughly removed, which finally results in higher alkali absorbance values and fail to meet the qualification standard; and because the solid crude product and the liquid crude product are treated separately, the loss is more caused by adding treatment procedures, and the yield is still about 3 percent lower than that of the method after the feed liquid is used mechanically.
Comparative examples 3-5 were slightly different from the examples in the oxidation, dilution, decolorization processes, but the quality of the finished product was poor; it can be seen that the oxidation, dilution, decolorization steps must be performed in the process sequence of the present invention.
The generic principles defined herein may be applied to 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 (1)
1. A subsequent treatment method for synthesizing D-p-hydroxyphenylglycine by an enzymatic method is characterized in that the method is a method for performing subsequent treatment on a feed liquid obtained by a reaction of p-hydroxyphenylglycine prepared by converting raw materials of p-hydroxyphenylglycine by using D-hydantoinase and D-carbamoyl hydrolase;
the method specifically comprises the following steps:
(1) Oxidation
Adjusting the pH value of a feed liquid obtained by reacting the D-p-hydroxyphenylglycine synthesized by an enzyme method to be alkaline, and adding hydrogen peroxide for oxidation to obtain an oxidized feed liquid;
(2) Dilution of
Adding water into the oxidation feed liquid for dilution to obtain D-p-hydroxyphenylglycine diluent;
(3) Decoloring (decoloring)
Adjusting the pH value of the D-p-hydroxyphenylglycine diluent to be acidic, adding activated carbon for adsorption and impurity removal, and filtering the activated carbon to obtain D-p-hydroxyphenylglycine decolorized solution;
(4) Obtaining the finished product
The D-p-hydroxyphenylglycine decolorization liquid is subjected to reverse osmosis concentration to separate out D-p-hydroxyphenylglycine, and the D-p-hydroxyphenylglycine decolorization liquid is filtered, washed and dried to obtain a finished product D-p-hydroxyphenylglycine;
in the step (1), the pH is adjusted to 10.0-10.2; the alkali used for adjusting the pH is the same as the alkali used for the reaction of synthesizing the D-p-hydroxyphenylglycine by an enzyme method; hydrogen peroxide is added until the feed liquid is not discolored;
in the step (2), the oxidation feed liquid is diluted 3-6 times;
in the step (3), the pH is adjusted to 5.0-5.2; the acid used for adjusting the pH is hydrochloric acid with the mass fraction of 20-30%; the dosage of the activated carbon is 1-5kg/m 3 D-p-hydroxyphenylglycine diluent, and the adsorption time is 20-60min;
in the step (4), a reducing agent is added into the D-p-hydroxyphenylglycine decolorization solution before reverse osmosis concentration to neutralize redundant hydrogen peroxide; reverse osmosis concentration multiple is 10-20 times;
further comprises:
(5) Monovalent salt recovery
Nanofiltration is carried out on the filtrate obtained by filtration in the step (4) to obtain a monovalent salt-containing dialysate and monovalent salt-removing liquid;
concentrating and evaporating the monovalent salt-containing dialysate to dryness to obtain monovalent salt;
(6) Trivalent salt recovery
Electrodialysis is carried out on the monovalent salt removal liquid obtained in the step (5) to obtain trivalent salt water and trivalent salt removal liquid; concentrating and evaporating trivalent salt brine to dryness to obtain trivalent salt;
the RO dialysate obtained by reverse osmosis concentration is returned to the step (2) for diluting the oxidation liquid or for nanofiltration; the 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 concentrating and evaporating is returned to the step (2) for diluting the oxidation feed liquid;
in the step (6), the trivalent salt removal liquid returns to the step (2) for diluting the oxidation liquid.
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