A method for penicillin fermentation extraction and its application

Abstract

The present invention relates to an efficient penicillin fermentation extraction method and application, comprising: performing coarse filtration, a two-stage ultrafiltration process, continuous extraction, purification, alkalinization dilution, and crystallization on the penicillin fermentation broth; the two-stage ultrafiltration process includes a first-stage ultrafiltration and a second-stage ultrafiltration; the first-stage ultrafiltration is to concentrate the concentrated liquid to a transmittance of 5-10%; the second-stage ultrafiltration is to concentrate the concentrated liquid obtained by the first-stage ultrafiltration to a transmittance of less than 1%; the continuous extraction comprises: combining the dialysates obtained by the first-stage ultrafiltration and the second-stage ultrafiltration to form a two-stage membrane dialysate; and mixing the two-stage membrane dialysate, butyl acetate, and dilute sulfuric acid by flow addition, and simultaneously extracting and centrifuging. The present invention utilizes a two-stage ultrafiltration and continuous extraction process to achieve continuous penicillin production, effectively reducing impurity content while improving production efficiency and penicillin yield. Furthermore, the amount of demulsifier and activated carbon used can be reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of biopharmaceuticals, and in particular to an efficient penicillin fermentation extraction method and application. Background Art

[0002] Large-scale production of penicillin typically utilizes fermentation. The resulting fermentation broth is complex, containing not only penicillin but also mycelium, pigments, amino acids, proteins, penicillin homologues, and degradation products. Penicillin isolation and purification involves filtration, extraction, and azeotropic crystallization. Traditional penicillin fermentation extraction methods suffer from significant penicillin loss, low production efficiency, high production costs, and poor penicillin product quality.

[0003] At present, the traditional filtration process can only roughly separate solid substances such as mycelium and large suspended matter in the fermentation broth, but cannot remove small molecules such as soluble proteins and miscellaneous sugars that exist in large quantities in the fermentation broth. The presence of these small molecule impurities and proteins is the root cause of the emulsification phenomenon. At the same time, it will greatly increase the load of subsequent processes such as solvent extraction, decolorization, evaporation and crystallization, thereby leading to large wastewater discharge, increased energy consumption, high production costs, unstable product quality and other problems.

[0004] Chinese patent CN1055290C optimizes the extraction process to reduce emulsification and improve yield. Chinese patent CN1500788A uses microfiltration to separate and extract penicillin. This method does not achieve continuous extraction of penicillin and does not completely eliminate the use of demulsifiers. Chinese patent CN1055291C uses an alcohol extractant instead of butyl acetate to improve the extraction rate, but still does not achieve continuous extraction of penicillin and does not improve production efficiency. Summary of the Invention

[0005] In order to solve the problems existing in the prior art, the present invention provides an efficient penicillin fermentation extraction method and application. By adopting a two-stage ultrafiltration and continuous extraction process, the continuous production of penicillin is achieved, which significantly improves the production efficiency and the penicillin yield.

[0006] The present invention provides a penicillin fermentation extraction method, comprising:

[0007] For penicillin fermentation broth, coarse filtration, two-stage ultrafiltration process, continuous extraction, purification, alkalinization dilution and crystallization are carried out;

[0008] The two-stage ultrafiltration process includes a first-stage ultrafiltration and a second-stage ultrafiltration;

[0009] The first-stage ultrafiltration is to concentrate the concentrated liquid until the transmittance is 5-10%; the second-stage ultrafiltration is to concentrate the concentrated liquid obtained by the first-stage ultrafiltration until the transmittance is less than 1%;

[0010] The continuous extraction comprises:

[0011] The dialysates obtained from the primary ultrafiltration and the secondary ultrafiltration are combined into a two-stage membrane dialysate; the two-stage membrane dialysate, butyl acetate and dilute sulfuric acid are added and mixed, and simultaneously subjected to extraction and centrifugation.

[0012] The present invention creatively adopts a process different from conventional continuous extraction. Conventional continuous extraction is generally continuous extraction in terms of solvent use (after evaporation, reflux continues to extract). The present invention is a continuous extraction in the overall process. After the extract, solvent and dilute sulfuric acid are added and mixed at the same time, they are directly introduced into the extraction centrifuge. The extraction effect is achieved in the pipeline. The extraction centrifuge is in a state of continuous operation during the entire process. The extracted solution is directly centrifuged and phase-separated, and the extract is discharged for subsequent processes. When using an extraction centrifuge in the prior art, extraction centrifugation is usually carried out in batches. The initial solution is mixed and then added to the extraction centrifuge. After adjusting the pH, extraction centrifugation is carried out; the next batch of extraction is started after the previous batch is completed. In comparison, the continuous extraction process adopted by the present invention can realize the continuous production of penicillin, significantly improve the extraction efficiency, and thus greatly improve the production efficiency.

[0013] In fact, in this process, the two-stage ultrafiltration process also plays a key role. It effectively intercepts mycelium, pigments, large molecular proteins, soluble proteins and other substances in the fermentation broth, making the filtrate high in light transmittance and low in impurities, thus avoiding the occurrence of emulsification in the subsequent extraction process. The filtrate obtained in this way can be mixed in the pipeline during the continuous flow addition process to quickly achieve the extraction effect, and then the extraction process can be carried out uninterruptedly.

[0014] Furthermore, the first-stage ultrafiltration adopts spiral membrane ultrafiltration, and the second-stage ultrafiltration adopts tubular membrane ultrafiltration.

[0015] The present invention adopts a spiral membrane ultrafiltration in the primary ultrafiltration process. This type of membrane is suitable for concentrating a feed liquid with relatively less suspended matter. When the concentration multiple is large, the membrane is easily blocked and not resistant to dirt. Tubular membrane ultrafiltration is used in the secondary ultrafiltration because although this type of membrane is not easy to block, the unit membrane area is small and the cost is high. Therefore, a spiral membrane is first used for ultrafiltration to concentrate to the stage where the membrane is prone to blockage, and then the concentrated liquid is ultrafiltered using a tubular membrane. In this way, the combination of the two ultrafiltration membranes can avoid the problems of membrane blockage and high production cost caused by using only a single membrane for ultrafiltration to the greatest extent.

[0016] Furthermore, the conditions of the first-stage ultrafiltration include: molecular weight 10000-30000 KDa, temperature 10-15 ° C, pressure 3-6 kg / cm 2 ;

[0017] The conditions of the secondary ultrafiltration include: molecular weight 10000-30000 KDa, temperature 10-15 ° C, pressure 3-5 kg / cm 2 .

[0018] Further, the continuous extraction comprises:

[0019] The two-stage membrane dialyzate, butyl acetate and dilute sulfuric acid are continuously added and mixed, passed through a high-efficiency mixer, and after being uniformly mixed in the high-efficiency mixer, directly enter an extraction centrifuge for centrifugal separation to obtain an extract.

[0020] Furthermore, the flow rate of the two-stage membrane dialyzate in the continuous extraction is 8 to 18 m 3 / h; the flow rate of butyl acetate in the continuous extraction is 6 to 10m 3 / h; the flow rate of the dilute sulfuric acid in the extraction centrifuge is 0.5 to 1.1m 3 / h, the pH of the mixed phase is 1.80-2.00.

[0021] Furthermore, the coarse filtration includes filtering the penicillin fermentation broth under conditions of a pore size of 5 to 15 μm and a pressure of ≤0.5 MPa.

[0022] Furthermore, the purification includes:

[0023] The extract obtained by the continuous extraction is treated with activated carbon at 0-10°C for 30-60 minutes and then washed with water; the flow rate of drinking water during the washing process is 5.0-8.0m 3 / h, total flow of decolorizing liquid and drinking water 20.0~35.0m 3 / h、V 脱色液 :V 饮用水 =3:1~5:1; the amount of the activated carbon is 10~30g / billion.

[0024] Furthermore, the alkalinization dilution comprises: adjusting the pH of the water washing liquid to 6 to 8 with a potassium carbonate solution, and then adding an alcohol ester mixture to dilute it;

[0025] The crystallization comprises: performing azeotropic crystallization under the condition of a gas phase temperature of 25 to 60°C.

[0026] As a preferred embodiment, the present invention provides a method for extracting penicillin by fermentation, comprising:

[0027] For penicillin fermentation broth, coarse filtration, two-stage ultrafiltration process, continuous extraction, purification, alkalinization dilution and crystallization are carried out;

[0028] The coarse filtration comprises: filtering the penicillin fermentation broth under conditions of a pore size of 5 to 15 μm and a pressure of ≤0.5 MPa;

[0029] The two-stage ultrafiltration process includes a first-stage ultrafiltration and a second-stage ultrafiltration;

[0030] The first-stage ultrafiltration is to use a spiral membrane to concentrate the concentrated liquid until the transmittance is 5-10%; the second-stage ultrafiltration is to use a tubular membrane to concentrate the concentrated liquid obtained by the first-stage ultrafiltration until the transmittance is less than 1%;

[0031] The conditions of the first-stage ultrafiltration include: molecular weight of 10,000 to 30,000 KDa, temperature of 10 to 15°C, pressure of 3 to 6 kg / cm 2 ;

[0032] The conditions of the secondary ultrafiltration include: molecular weight 10000-30000 KDa, temperature 10-15 ° C, pressure 3-5 kg / cm 2 ;

[0033] The continuous extraction comprises:

[0034] The two-stage membrane dialysate, butyl acetate and dilute sulfuric acid are continuously added and mixed, and then passed through a high-efficiency mixer. After being evenly mixed in the high-efficiency mixer, they are directly fed into an extraction centrifuge for centrifugal separation to obtain an extract; the flow rate of the two-stage membrane dialysate in the continuous extraction is 8 to 18 m 3 / h; the flow rate of butyl acetate in the continuous extraction is 6 to 10m 3 / h; the flow rate of the dilute sulfuric acid in the extraction centrifuge is 0.5 to 1.1m 3 / h; the pH of the mixed phase is 1.80-2.00.

[0035] The purification comprises:

[0036] The extract is treated with activated carbon at 0-10°C for 30-60 minutes and then washed with water; the flow rate of drinking water during the washing process is 5.0-8.0 m 3 / h, total flow of decolorizing liquid and drinking water 20.0~35.0m 3 / h、V 脱色液 :V 饮用水 =3:1~5:1; the amount of activated carbon used is 10~30g / billion;

[0037] The alkalization dilution comprises:

[0038] After adjusting the pH of the water wash to 6-8 with potassium carbonate solution, add alcohol ester mixture to dilute;

[0039] The crystallization comprises:

[0040] Azeotropic crystallization is carried out at a vapor phase temperature of 40 to 45°C.

[0041] Furthermore, the flow rate of the water washing liquid during the alkalinization dilution process is 15 to 30 m 3 / h, the flow rate of potassium carbonate is 0.5~1.1m 3 / h.

[0042] The present invention further provides application of the penicillin fermentation extraction method in improving penicillin yield or production efficiency.

[0043] The present invention has the following beneficial effects:

[0044] The present invention provides an efficient penicillin fermentation extraction method and application, which realizes the continuous production of penicillin through a two-stage ultrafiltration and continuous extraction process, effectively removes mycelium, pigments, macromolecular proteins, soluble proteins and other substances in the fermentation broth, and improves production efficiency and penicillin yield.

[0045] The penicillin fermentation extraction method provided by the present invention has an extraction yield of penicillin exceeding 98%. Due to the adoption of a two-stage ultrafiltration process + continuous extraction technical means, the occurrence of emulsification is avoided during the extraction process, the use of a demulsifier is eliminated, and the amount of activated carbon used is reduced, thereby improving production efficiency, reducing production costs, and enhancing the quality of subsequent penicillin products. At the same time, the difficulty of treating subsequent penicillin waste acid water is reduced, thereby further reducing production costs. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the technical solutions in the present invention or the prior art, a brief introduction is given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0047] Figure 1 This is a flow chart of the efficient penicillin fermentation extraction method provided in Example 1 of the present invention. DETAILED DESCRIPTION

[0048] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0049] The conditions in the following embodiments are expressed in the form of ranges because it is difficult to maintain a fixed condition in actual production practice. Therefore, it is sufficient to maintain it within a certain range in the steps.

[0050] Example 1

[0051] This embodiment provides an efficient penicillin fermentation extraction method, such as Figure 1 The specific process is as follows:

[0052] Fermentation broth plate and frame filtration: 5100L of penicillin fermentation broth was filtered through a plate and frame filter press with a filter cloth pore size of 10μm and a filtration pressure of 0.5MPa. Mycelium, large suspended matter and other solids in the fermentation broth were removed to obtain 2900L of filtrate with a titer of 61022ug / mL and a transmittance of 16.96%.

[0053] Filtrate spiral membrane ultrafiltration: After the spiral membrane is cleaned and qualified, the membrane equipment is opened to ultrafilter 2900L of filtrate. The membrane molecular weight cutoff is 30000, the ultrafiltration temperature is 12-14℃, and the operating pressure is 4-4.5kg / cm 2 In the early stage, full reflux filtration was used. After the dialysate was clarified, the discharge was switched until the ultrafiltration was completed. About 2705L of dialysate and about 190L of concentrated liquid were discharged. The transmittance of the dialysate was 78.19%, and the transmittance of the concentrated liquid was 8.2%.

[0054] Tubular membrane ultrafiltration of concentrated liquid: After the tubular membrane is cleaned and qualified, the membrane equipment is opened to re-filter 190L of concentrated liquid. The membrane molecular weight cut-off is 30,000, the ultrafiltration temperature is 13-14°C, and the pressure before the membrane is 3.5-4.0 kg / cm 2 , pressure behind the membrane 2.0~2.5kg / cm 2 In the early stage, full reflux filtration was used, and the discharge was switched after the dialysate was clarified. 295L of water was added during the operation until the penicillin potency of the retentate was 680μg / mL. The concentration was continued until the flux was greatly reduced. The dialysate output was about 435L, the retentate was about 45L, the dialysate transmittance was 71.36%, and the concentrate transmittance was 0.22%.

[0055] Continuous extraction and separation of dialysate: The combined two-stage membrane dialysate was approximately 3140 L, with a titer of 55370 μg / mL and a transmittance of 72.33%. The extraction centrifuge was started, and the regulating valve controlled the dialysate flow rate to 12.0 m 3 / h, butyl acetate flow rate 8.0m 3 / h, dilute sulfuric acid flow rate 0.7m 3 / h, the three solutions were continuously passed through a high-efficiency mixer for rapid mixing (in this case, the pH was maintained at 1.80-2.00), and then continuously passed through an extraction centrifuge for centrifugal separation to obtain 2300 L of light phase extract BA with a titer of 74960 μg / mL and an extraction yield of 99.16%.

[0056] Penicillin purification: 2300L of extract BA was pumped into the decolorization tank, 2.59Kg of activated carbon (15g / 1 billion) was added, and the decolorization was carried out at 5-8℃ with stirring for 45 minutes. The carbon was removed by filtering with a self-cleaning filter to obtain the decolorized liquid FBA. The total flow rate of the decolorized liquid and drinking water was adjusted to 20m 3 / h, drinking water flow rate 5m 3 / h, control the ratio of decolorizing liquid to drinking water to be 3:1, pump 2300L of decolorizing liquid and drinking water into the water washing centrifuge through the pipeline, centrifuge and separate the phases, and the light phase obtained is the water washing liquid RBA.

[0057] Alkalization and dilution: adjust the flow rate of water washing liquid RBA to 20m 3 / h, potassium carbonate flow rate 0.8m 3 / h, 2300L of water washing liquid and potassium carbonate are mixed through a pipeline, the pH of the mixed liquid is controlled at 6.35-6.80, and the mixed liquid enters a mixed liquid tank, is stirred, and pumped into an alkalization liquid tank, and allowed to stand for phase separation to obtain 198L of lower alkalization liquid with a titer of 869643ug / mL; 198L of alkalization liquid is pumped into a dilution tank, 293L of alcohol-ester mixture is added and mixed to obtain 491L of diluted liquid with a titer of 350200ug / mL.

[0058] Azeotropic crystallization: Pump 491L of diluent into the crystallizer. Control the stirring frequency at 40.00Hz and the vacuum at ≥0.080MPa. Open the steam valve and control the vapor phase temperature at 41.0-43.0°C for crystallization. When the liquid begins to boil, add additional alcohol-ester mixture, maintaining a stable liquid level. When crystals appear in the tank, reduce the stirring frequency by 25.00Hz, close the steam valve, stop adding the alcohol-ester mixture, and grow the crystals for 45 minutes. Upon completion, adjust the stirring frequency to 35.00Hz, open the steam valve, control the vapor phase temperature at 43.0-45.0°C, and continue crystallization, maintaining the same amount of alcohol-ester mixture in both input and output. Distill until the mother liquor moisture content reaches 0.91%, then stop distillation and cool for 15 minutes. Place the crystallizer in a three-in-one tank and filter. Rinse the wet powder with 150L of alcohol-ester mixture for 20 minutes and drain. Maintain a vacuum at ≥0.080MPa and a vapor phase temperature at 50°C, and dry for 180 minutes. After drying, the product was weighed and packaged to obtain 104.61 kg of finished product of potassium penicillin industrial salt with a yield of 95.04%.

[0059] Example 2

[0060] This embodiment provides an efficient penicillin fermentation extraction method, the specific process is as follows:

[0061] Fermentation broth plate and frame filtration: 5000L of penicillin fermentation broth was filtered through a plate and frame filter press with a filter cloth pore size of 10μm and a filtration pressure of 0.5MPa. Mycelium, large suspended matter and other solid substances in the fermentation broth were removed to obtain 2950L of filtrate with a titer of 59980ug / mL and a transmittance of 17.50%.

[0062] Filtrate spiral membrane ultrafiltration: After the spiral membrane is cleaned and qualified, the membrane equipment is opened to ultrafilter 2950L of filtrate, with a membrane molecular weight cutoff of 30,000, an ultrafiltration temperature of 13-14°C, and an operating pressure of 4-4.5kg / cm 2 In the early stage, full reflux filtration was used. After the dialysate was clarified, the discharge was switched until the ultrafiltration was completed. About 2740L of dialysate and about 200L of concentrated liquid were discharged. The transmittance of the dialysate was 80.26%, and the transmittance of the concentrated liquid was 6.6%.

[0063] Tubular membrane ultrafiltration of concentrated liquid: After the tubular membrane is cleaned and qualified, the membrane equipment is opened to re-filter 200L of concentrated liquid. The membrane molecular weight cut-off is 30,000, the ultrafiltration temperature is 13-14°C, and the pressure before the membrane is 3.0-3.5kg / cm 2 , pressure behind the membrane 2.5~3.0kg / cm 2 In the early stage, full reflux filtration was used. After the dialysate was clarified, the discharge was switched. During the operation, 310L of water was added until the penicillin potency of the retentate was 725μg / mL. The concentration was continued until the flux was greatly reduced. The dialysate output was about 460L, the retentate was about 48L, the dialysate transmittance was 74.22%, and the concentrate transmittance was 0.36%.

[0064] Continuous extraction of dialysate: The combined two-stage membrane dialysate was approximately 3200 L, with a titer of 53890 μg / mL and a transmittance of 76.84%. The extraction centrifuge was started, and the regulating valve controlled the dialysate flow rate at 12.0 m 3 / h, butyl acetate flow rate 8.0m 3 / h, dilute sulfuric acid flow rate 0.7m 3 / h, the three solutions were continuously passed through a high-efficiency mixer for rapid mixing (in this case, the pH was maintained at 1.80-2.00), and then continuously passed through an extraction centrifuge for centrifugal separation to obtain 2190 L of light phase extract BA, with a titer of 77483 ug / mL and an extraction yield of 98.40%.

[0065] Penicillin purification: 2190L of extract BA was pumped into the decolorization tank, 2.55Kg of activated carbon (15g / 1 billion) was added, and the decolorization was carried out under stirring at 8-10℃ for 45 minutes. The carbon was removed by filtering with a self-cleaning filter to obtain the decolorized liquid FBA. The total flow rate of the decolorized liquid and drinking water was adjusted to 20m 3 / h, drinking water flow rate 5m 3 / h, control the ratio of decolorizing liquid to drinking water at 3:1, pump 2190L of decolorizing liquid and drinking water into the water washing centrifuge through the pipeline, centrifuge and separate the phases, and the light phase obtained is the water washing liquid RBA.

[0066] Alkalization and dilution: adjust the flow rate of water washing liquid RBA to 20m 3 / h, potassium carbonate flow rate 0.8m3 / h, 2190L of water washing liquid and potassium carbonate are mixed through a pipeline, the pH of the mixed liquid is controlled at 6.40-6.75, and the mixed liquid enters a mixed liquid tank, is stirred, and pumped into an alkalization liquid tank, and allowed to stand for phase separation to obtain 203L of lower alkalization liquid with a titer of 850930ug / mL; 203L of alkalization liquid is pumped into a dilution tank, 274L of alcohol-ester mixture is added and mixed to obtain 477L of diluted liquid with a titer of 362154ug / mL.

[0067] Azeotropic crystallization: Pump 477L of diluent into the crystallizer. Control the stirring frequency at 40.00Hz and the vacuum at ≥0.080MPa. Open the steam valve and control the vapor phase temperature at 44.0-45.0°C for crystallization. When the liquid begins to boil, add additional alcohol-ester mixture, maintaining a stable liquid level. When crystals appear in the tank, reduce the stirring frequency by 25.00Hz, close the steam valve, stop adding the alcohol-ester mixture, and grow the crystals for 45 minutes. Upon completion, adjust the stirring frequency to 35.00Hz, open the steam valve, control the vapor phase temperature at 42.0-44.0°C, and continue crystallization, maintaining the same amount of alcohol-ester mixture in both input and output. Distill until the mother liquor moisture reaches 0.78%, then stop distillation and cool for 15 minutes. Place the crystallizer in a three-in-one tank and filter. Rinse the wet powder with 150L of alcohol-ester mixture for 20 minutes and drain. Maintain a vacuum at ≥0.080MPa and a vapor phase temperature at 45°C, and dry for 210 minutes. After drying, the product was weighed and packaged to obtain 106.01 kg of finished product of potassium penicillin industrial salt with a yield of 95.86%.

[0068] Comparative Example 1

[0069] This comparative example provides a traditional penicillin fermentation extraction method, the specific process is as follows:

[0070] Fermentation broth plate and frame filtration: 5000L of penicillin fermentation broth was filtered through a plate and frame filter press with a filter cloth pore size of 10μm and a filtration pressure of 0.5MPa. Mycelium, large suspended matter and other solids in the fermentation broth were removed to obtain 2800L of filtrate with a titer of 58174ug / mL and a transmittance of 17.68%.

[0071] Filtrate extraction: 2800L of filtrate was pumped into the extraction tank, 2100L of butyl acetate and 280L of 2% demulsifier were added, and dilute sulfuric acid was added to adjust the pH to 1.80-2.00. The mixture was allowed to stand for phase separation, and the aqueous phase was taken to the recovery station to recover butyl acetate. The organic phase was the extract BA, with a titer of 74430ug / mL and an extraction yield of 95.96%.

[0072] Penicillin purification: 2100L of extract BA was pumped into the decolorization tank, 9kg of activated carbon (60g / 1 billion) was added, and the mixture was stirred and decolorized at 6-8℃ for 45min. The carbon was removed by filtering with a self-cleaning filter to obtain the decolorized liquid FBA. The total flow rate of the decolorized liquid and drinking water was adjusted to 20m 3 / h, drinking water flow rate 5m 3 / h, control the ratio of decolorizing liquid to drinking water to be 3:1, pump 2100L of decolorizing liquid and drinking water into the water washing centrifuge through the pipeline, centrifuge and separate the phases, and the light phase obtained is the water washing liquid RBA.

[0073] Alkalization and dilution: adjust the flow rate of water washing liquid RBA to 20m 3 / h, potassium carbonate flow rate 0.8m 3 / h, 2100L of water washing liquid and potassium carbonate are mixed through a pipeline, the pH of the mixed liquid is controlled at 6.30-6.55, and the mixed liquid enters a mixed liquid tank, is stirred, and pumped into an alkalization liquid tank, and allowed to stand for phase separation to obtain 181L of lower alkalization liquid with a titer of 831699ug / mL; 181L of alkalization liquid is pumped into a dilution tank, 240L of alcohol-ester mixture is added and mixed to obtain 421L of diluted liquid with a titer of 357362ug / mL.

[0074] Azeotropic crystallization: Pump 421L of diluent into the crystallizer. Control the stirring frequency at 40.00Hz and the vacuum at ≥0.080MPa. Open the steam valve and control the vapor phase temperature at 42.0-44.0°C for crystallization. When the liquid begins to boil, add additional alcohol-ester mixture, maintaining a stable liquid level. When crystals appear in the tank, reduce the stirring frequency by 25.00Hz, close the steam valve, stop adding the alcohol-ester mixture, and grow the crystals for 45 minutes. Upon completion, adjust the stirring frequency to 35.00Hz, open the steam valve, control the vapor phase temperature at 44.0-45.0°C, and continue crystallization, maintaining the same amount of alcohol-ester mixture in both input and output. Distill until the mother liquor moisture reaches 0.96%, then stop distillation and cool for 15 minutes. Place the crystallized liquid into a three-in-one tank and filter. Rinse the wet powder with 150L of alcohol-ester mixture for 20 minutes and drain. Maintain a vacuum at ≥0.080MPa and a vapor phase temperature at 50°C, and dry for 180 minutes. After drying, the product was weighed and packaged to obtain 90.78 kg of finished product of potassium penicillin industrial salt with a yield of 94.26%.

[0075] Comparative Example 2

[0076] This comparative example provides a traditional penicillin fermentation extraction method, the specific process is as follows:

[0077] Fermentation broth plate and frame filtration: 5500L of penicillin fermentation broth was filtered through a plate and frame filter press with a filter cloth pore size of 10μm and a filtration pressure of 0.5MPa. Mycelium, large suspended matter and other solids in the fermentation broth were removed to obtain 3100L of filtrate with a titer of 60188ug / mL and a transmittance of 19.25%.

[0078] Filtrate extraction: 3100L of filtrate was pumped into the extraction tank, 2500L of butyl acetate and 310L of 2% demulsifier were added, and dilute sulfuric acid was added to adjust the pH to 1.80-2.00. The mixture was allowed to stand for phase separation, and the aqueous phase was taken to the recovery station to recover butyl acetate. The organic phase was the extract BA, with a titer of 71695ug / mL and an extraction yield of 96.06%.

[0079] Penicillin purification: 2500L of extract BA was pumped into the decolorization tank, 10.5Kg of activated carbon (60g / 1 billion) was added, and the decolorization was carried out at 4-6℃ with stirring for 45 minutes. The carbon was removed by filtering with a self-cleaning filter to obtain the decolorized liquid FBA. The total flow rate of the decolorized liquid and drinking water was adjusted to 20m 3 / h, drinking water flow rate 5m 3 / h, control the ratio of decolorizing liquid to drinking water to be 3:1, pump 2500L of decolorizing liquid and drinking water into the water washing centrifuge through the pipeline, centrifuge and separate the phases, and the light phase obtained is the water washing liquid RBA.

[0080] Alkalization and dilution: adjust the flow rate of water washing liquid RBA to 20m 3 / h, potassium carbonate flow rate 0.8m 3 / h, 2500L of water washing liquid and potassium carbonate are mixed through a pipeline, the pH of the mixed liquid is controlled at 6.45-6.75, and the mixed liquid enters a mixed liquid tank, is stirred, and pumped into an alkalization liquid tank, and allowed to stand for phase separation to obtain 213L of lower alkalization liquid with a titer of 821376ug / mL; 213L of alkalization liquid is pumped into a dilution tank, 265L of alcohol-ester mixture is added and mixed to obtain 478L of diluted liquid with a titer of 365842ug / mL.

[0081] Azeotropic crystallization: Pump 478L of diluent into the crystallizer. Control the stirring frequency at 40.00Hz and the vacuum at ≥0.080MPa. Open the steam valve and control the vapor phase temperature at 40.0-42.0°C for crystallization. When the liquid begins to boil, add additional alcohol-ester mixture, maintaining a stable liquid level. When crystals appear in the tank, reduce the stirring frequency by 25.00Hz, close the steam valve, stop adding the alcohol-ester mixture, and grow the crystals for 45 minutes. Upon completion, adjust the stirring frequency to 35.00Hz, open the steam valve, control the vapor phase temperature at 42.0-43.0°C, and continue crystallization, maintaining the same amount of alcohol-ester mixture in both input and output. Distill until the mother liquor moisture content reaches 0.82%, then stop distillation and cold pump for 15 minutes. Place the crystallized liquid into a three-in-one tank and filter. Rinse the wet powder with 150L of alcohol-ester mixture for 20 minutes and drain. Maintain a vacuum at ≥0.080MPa and a vapor phase temperature at 45°C, and dry for 210 minutes. After drying, the product was weighed and packaged to obtain 105.11 kg of finished product of potassium penicillin industrial salt with a yield of 93.89%.

[0082] Table 1 Summary of test data of the above embodiment

[0083]

[0084]

[0085] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A penicillin fermentation extraction method, characterized in that: include: For penicillin fermentation broth, coarse filtration, two-stage ultrafiltration process, continuous extraction, purification, alkalinization dilution and crystallization are carried out; The two-stage ultrafiltration process includes a first-stage ultrafiltration and a second-stage ultrafiltration; The first stage ultrafiltration is concentrated to a concentrated liquid with a transmittance of 5-10%; The secondary ultrafiltration is to concentrate the concentrated solution obtained by the primary ultrafiltration until the transmittance is less than 1%; The continuous extraction comprises: The dialysates obtained from the first-stage ultrafiltration and the second-stage ultrafiltration are combined to form a two-stage membrane dialysate; the two-stage membrane dialysate, butyl acetate, and dilute sulfuric acid are continuously added and mixed, and passed through a high-efficiency mixer. After being uniformly mixed in the high-efficiency mixer, the mixture is directly fed into an extraction centrifuge for centrifugal separation to obtain an extract; The first-stage ultrafiltration adopts spiral membrane ultrafiltration, and the second-stage ultrafiltration adopts tubular membrane ultrafiltration; the conditions of the first-stage ultrafiltration include: molecular weight 10000~30000KDa, temperature 10~15℃, pressure 3~6 kg / cm 2 ; The conditions of the secondary ultrafiltration include: molecular weight 10000~30000KDa, temperature 10~15℃, pressure 3~5 kg / cm 2 ; The flow rate of the two-stage membrane dialysate in the continuous extraction is 8~18 m 3 / h; the flow rate of butyl acetate in the continuous extraction is 6~10 m 3 / h; the flow rate of the dilute sulfuric acid in the continuous extraction is 0.5~1.1 m 3 / h, the pH of the mixed phase is 1.80~2.

00.

2. The penicillin fermentation extraction method according to claim 1, characterized in that: The coarse filtration comprises filtering the penicillin fermentation broth under the conditions of a pore size of 5 to 15 μm and a pressure of ≤0.5 MPa.

3. The penicillin fermentation extraction method according to any one of claims 1 to 2, characterized in that: The purification comprises: The extract obtained by the continuous extraction is treated with activated carbon at 0-10°C for 30-60 minutes and then washed with water; the drinking water flow rate during the washing process is 5.0-8.0m 3 / h, total flow of decolorizing liquid and drinking water 20.0~35.0m 3 / h、V 脱色液 :V 饮用水 =3:1~5:1; the amount of the activated carbon is 10~30g / billion.

4. The penicillin fermentation extraction method according to any one of claims 1 to 2, characterized in that: The alkalinization dilution comprises: adjusting the pH of the water washing liquid to 6-8 with potassium carbonate solution, and then adding an alcohol ester mixture to dilute; The crystallization comprises: performing azeotropic crystallization under the condition of a gas phase temperature of 25-60°C.

5. The penicillin fermentation extraction method according to claim 1, characterized in that: include: For penicillin fermentation broth, coarse filtration, two-stage ultrafiltration process, continuous extraction, purification, alkalinization dilution and crystallization are carried out; The coarse filtration comprises: filtering the penicillin fermentation broth under conditions of a pore size of 5 to 15 μm and a pressure of ≤0.5 MPa; The two-stage ultrafiltration process includes a first-stage ultrafiltration and a second-stage ultrafiltration; The first-stage ultrafiltration is to use a spiral membrane to concentrate the concentrated liquid until the transmittance is 5-10%; the second-stage ultrafiltration is to use a tubular membrane to concentrate the concentrated liquid obtained by the first-stage ultrafiltration until the transmittance is less than 1%; The conditions of the first-stage ultrafiltration include: molecular weight 10000~30000KDa, temperature 10~15℃, pressure 3~6 kg / cm 2 ; The conditions of the secondary ultrafiltration include: molecular weight 10000~30000KDa, temperature 10~15℃, pressure 3~5 kg / cm 2 ; The continuous extraction comprises: The two-stage membrane dialysate, butyl acetate and dilute sulfuric acid are continuously added and mixed, and then passed through a high-efficiency mixer. After being evenly mixed in the high-efficiency mixer, they are directly fed into an extraction centrifuge for centrifugal separation to obtain an extract; the flow rate of the two-stage membrane dialysate in the continuous extraction is 8-18 m 3 / h; the flow rate of butyl acetate in the continuous extraction is 6~10 m 3 / h; the flow rate of the dilute sulfuric acid in the extraction centrifuge is 0.5~1.1 m 3 / h, the pH of the mixed phase is 1.80~2.00; The purification comprises: The extract is treated with activated carbon at 0-10°C for 30-60 minutes and then washed with water; the drinking water flow rate during the washing process is 5.0-8.0 m 3 / h, total flow of decolorizing liquid and drinking water 20.0~35.0m 3 / h、V 脱色液 :V 饮用水 =3:1~5:1; the amount of activated carbon is 10~30g / billion; The alkalization dilution comprises: After adjusting the pH of the water wash to 6-8 with potassium carbonate solution, add alcohol ester mixture to dilute; The crystallization comprises: Azeotropic crystallization is carried out at a vapor phase temperature of 40-45°C.

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