CN109836346B - Purification process of isoleucine fermentation liquor - Google Patents
Purification process of isoleucine fermentation liquor Download PDFInfo
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Abstract
The invention provides a purification process of isoleucine fermentation broth, belonging to the technical field of isoleucine purification. Firstly, carrying out primary dialysis on isoleucine fermentation liquor by a ceramic membrane to remove most of thalli, impure proteins and colloids; then, the primary dialysate is subjected to secondary dialysis through a nanofiltration membrane to remove most of pigments, proteins and salts; carrying out continuous chromatographic separation on the secondary dialysate, and separating isoleucine from salt to obtain a product liquid with low impurity content; and performing tertiary dialysis on the obtained product liquid through a reverse osmosis membrane to obtain tertiary concentrated liquid with higher isoleucine content, and performing activated carbon decoloration, freeze crystallization and drying on the tertiary concentrated liquid to obtain isoleucine. The purification process provided by the invention does not need acid regulation and adjustment back to neutral treatment, the nanofiltration membrane plays a certain role in decolorization, the consumption of active carbon is low, continuous chromatographic separation is adopted, the utilization rate of resin is high, and the purification process also has the advantages of low wastewater discharge and low energy consumption.
Description
Technical Field
The invention relates to the technical field of isoleucine fermentation broth purification, in particular to a process for purifying isoleucine fermentation broth.
Background
Isoleucine is one of essential amino acids in the human body and plays an important role in the metabolism of life. Isoleucine is widely used in foods and medicines as a nutrient and a drug for treating diseases. Isoleucine is usually prepared by fermentation, the obtained fermentation liquor contains thalli, protein, sugar, salt, impurity amino acid, isoleucine and the like, and isoleucine can be obtained by purifying the fermentation liquor. In the prior art, the method for purifying the isoleucine fermentation liquor generally comprises the steps of firstly adding sulfuric acid to adjust the pH value of the fermentation liquor so as to coagulate protein, then removing coagulated impurities through a ceramic filter membrane, and then extracting isoleucine through ion exchange resin. In the purification process, a large amount of sulfuric acid is needed, new impurities are introduced, the utilization rate of the ion exchange resin is low, a regenerant is needed to be regenerated, and then washing is carried out, so that a large amount of wastewater is generated.
Disclosure of Invention
The invention aims to provide a process for purifying isoleucine fermentation broth, which does not need sulfuric acid and has less wastewater.
In order to achieve the above object, the present invention provides the following technical solutions:
a purification process of isoleucine fermentation broth comprises the following steps:
(1) performing primary dialysis on the isoleucine fermentation broth through a ceramic membrane to obtain primary dialysate and primary concentrated solution;
(2) performing secondary dialysis on the primary dialysate through a nanofiltration membrane to obtain secondary dialysate and secondary concentrated solution;
(3) carrying out continuous chromatographic separation on the secondary dialysate to obtain a product liquid and an impurity liquid;
(4) performing third-stage dialysis on the product liquid through a first reverse osmosis membrane to obtain a third-stage concentrated liquid and a third-stage dialysate;
(5) and sequentially carrying out activated carbon decoloration, freeze crystallization and drying on the third-level concentrated solution to obtain isoleucine.
Preferably, the molecular weight cut-off of the ceramic membrane is 45-55 KD; the pressure of the primary dialysis is 3-4 bar, and the temperature is 40-60 ℃.
Preferably, the molecular weight cut-off of the nanofiltration membrane is 450-550 Da; the pressure of the secondary dialysis is 10-18 bar, and the temperature is 48-55 ℃.
Preferably, the continuous chromatographic separation system comprises an elution zone, an extension separation zone, a main separation zone and a post separation zone, wherein each zone independently comprises 2-5 resin columns which are connected in series or in parallel.
Preferably, the rejection rate of the first reverse osmosis membrane to sodium chloride is more than or equal to 96 percent; the pressure of the three-stage dialysis is 12-16 bar, and the temperature is 40-52 ℃.
Preferably, the primary dialysis, the secondary dialysis and the tertiary dialysis are independently and precisely filtered, the filtrate is reserved for the primary dialysis, the secondary dialysis and the tertiary dialysis, the aperture of the precise filtering filter hole before the primary dialysis is 1-50 μm, and the aperture of the precise filtering filter hole before the secondary dialysis and the tertiary dialysis is independently 1-10 μm.
Preferably, after the first-stage dialysis is finished, the first-stage concentrated solution is subjected to first top washing, and water for the first top washing is a third-stage dialysate and/or an impurity solution filtered by a second reverse osmosis membrane; the volume of the first top washing water is 35-45% of the volume of the isoleucine fermentation broth.
Preferably, after the second-stage dialysis is finished, second top washing is carried out on the second-stage concentrated solution, and water for the second top washing is third-stage dialysate and/or impurity liquid filtered by a second reverse osmosis membrane; the volume of the second top washing water is 10-20% of the first-level dialysate.
Preferably, the mode of the primary dialysis, the secondary dialysis and the tertiary dialysis is cross-flow dialysis.
The invention provides a purification process of isoleucine fermentation broth, which comprises the following steps: (1) performing primary dialysis on the isoleucine fermentation broth through a ceramic membrane to obtain primary dialysate and primary concentrated solution; (2) performing secondary dialysis on the primary dialysate through a nanofiltration membrane to obtain secondary dialysate and secondary concentrated solution; (3) carrying out continuous chromatographic separation on the secondary dialysate to obtain a product liquid and an impurity liquid; (4) performing third-stage dialysis on the product liquid through a first reverse osmosis membrane to obtain a third-stage concentrated liquid and a third-stage dialysate; (5) and sequentially carrying out activated carbon decoloration, freeze crystallization and drying on the third-level concentrated solution to obtain isoleucine. Firstly, carrying out primary dialysis on isoleucine fermentation liquor by a ceramic membrane to remove most of thalli, impure proteins and colloids; then, performing secondary dialysis on the obtained primary dialysate through a nanofiltration membrane to remove most of pigments, proteins and salts; carrying out continuous chromatographic separation on the secondary dialysate, and separating isoleucine from salt to obtain a product liquid with low impurity content; and performing three-stage dialysis on the obtained product liquid through a reverse osmosis membrane, concentrating the product liquid to obtain a three-stage concentrated liquid with higher isoleucine content, and then performing activated carbon decoloration, freeze crystallization and drying on the three-stage concentrated liquid to obtain the isoleucine with higher purity.
The purification process provided by the invention does not need acid regulation and adjustment back to neutral, so that the dosage of acid and alkali is reduced, the nanofiltration membrane plays a certain role in decolorization, the dosage of active carbon is reduced, the utilization rate of resin is improved and the dosage of resin is reduced by using a continuous chromatographic separation method, and meanwhile, three-stage dialysate and impurity liquid in the purification process provided by the invention can be recycled, so that the discharge amount of wastewater is reduced, and the concentration of product liquid is realized by adopting a reverse osmosis membrane for three-stage dialysis, so that the purification process has the advantage of low energy consumption. Experimental results show that the yield of isoleucine by the purification process provided by the invention is over 88%, and the purity is 99.5-99.8%.
Drawings
FIG. 1 is a flow chart of the purification process of isoleucine fermentation broth provided by the present invention;
FIG. 2 is a flow chart of continuous chromatographic separation in the process for purifying isoleucine fermentation broth provided by the present invention.
Detailed Description
The invention provides a purification process of isoleucine fermentation broth, which comprises the following steps:
(1) performing primary dialysis on the isoleucine fermentation broth through a ceramic membrane to obtain primary dialysate and primary concentrated solution;
(2) performing secondary dialysis on the primary dialysate through a nanofiltration membrane to obtain secondary dialysate and secondary concentrated solution;
(3) carrying out continuous chromatographic separation on the secondary dialysate to obtain a product liquid and an impurity liquid;
(4) performing third-stage dialysis on the product liquid through a first reverse osmosis membrane to obtain a third-stage concentrated liquid and a third-stage dialysate;
(5) and sequentially carrying out activated carbon decoloration, freeze crystallization and drying on the third-level concentrated solution to obtain isoleucine.
FIG. 1 is a process flow diagram of the purification process of isoleucine fermentation broth provided by the present invention, wherein isoleucine fermentation broth is first dialyzed by ceramic membrane to remove most of thallus, foreign protein and colloid, and the obtained first-stage concentrated solution is subjected to environmental protection treatment; then, performing secondary dialysis on the primary dialysate through a nanofiltration membrane to remove most of pigments, proteins and salts, and performing environment-friendly treatment on the obtained secondary concentrated solution; carrying out continuous chromatographic separation on the secondary dialysate, and separating isoleucine from salt to obtain impurity liquid and product liquid with low impurity content, wherein the impurity liquid is preferably recycled after being filtered by a second reverse osmosis membrane; performing third-stage dialysis on the obtained product liquid through a first reverse osmosis membrane, and concentrating the product liquid to obtain third-stage dialysate and third-stage concentrated liquid with higher isoleucine content, wherein the third-stage dialysate can be directly recycled; and then the third-level concentrated solution is decolorized by active carbon, frozen, crystallized and dried to obtain the isoleucine with higher purity.
The method comprises the step of carrying out primary dialysis on isoleucine fermentation broth through a ceramic membrane to obtain primary dialysate and primary concentrated solution. In the present invention, the primary dialysis can remove most of the bacteria, impure proteins and colloids.
The physical and chemical properties of the isoleucine fermentation broth are not particularly limited, and the method is suitable for any isoleucine fermentation broth. In the embodiment of the invention, the isoleucine fermentation liquid is earthy yellow, the pH value is 6.5-6.9, the conductivity is 20000-25000 mu s/cm, and the content of isoleucine is 21-23 g/L.
The invention preferably carries out precise filtration before the first-stage dialysis, and the filtrate is left for the first-stage dialysis; the aperture of the filter hole for the precision filtration is preferably 1-50 μm; the operating conditions of the microfiltration are the same as for the first-stage dialysis. In the invention, the precise filtration has the function of filtering out large suspended matters and particles, thereby ensuring the safe and stable operation of the subsequent membrane process.
In the invention, the molecular weight cut-off of the ceramic membrane is preferably 45-55 KD, and more preferably 50 KD; the pressure of the primary dialysis is preferably 3-4 bar, the temperature is preferably 40-60 ℃, and more preferably 45-55 ℃. In the invention, after primary dialysis, the light transmittance of the isoleucine fermentation liquor is improved from 0.05% to about 11%, the protein removal rate can reach about 80%, and the concentration multiple of the concentrated solution obtained by the primary dialysis is 5-6; the average flux can reach 100-120 LMH.
In the invention, after the first-stage dialysis is finished, the first-stage concentrated solution is preferably subjected to first top washing, and the first top washing water is preferably a third-stage dialysate and/or an impurity solution filtered by a second reverse osmosis membrane; the volume of the first top washing water is preferably 35-45% of the volume of the isoleucine fermentation broth. In the present invention, it is preferable that the top washing solution obtained by the first top washing is dialyzed, and the obtained dialysate is combined with the dialysate obtained by the first-stage dialysis to be used as a first-stage dialysate, and the subsequent steps are performed; the dialysis conditions of the top wash solution were the same as the primary dialysis. In the invention, the recovery rate of isoleucine can be improved by top washing the primary concentrated solution; the washed first-stage concentrated solution is subjected to environment-friendly treatment (such as incineration and the like).
After the primary dialysate is obtained, the primary dialysate is subjected to secondary dialysis through a nanofiltration membrane to obtain secondary dialysate and secondary concentrated solution. In the present invention, the secondary dialysis removes most of the pigments, proteins and salts.
According to the method, the primary dialysate is preferably subjected to precise filtration before secondary dialysis, filtrate is reserved for primary dialysis, and the aperture of a filter hole of the precise filtration is preferably 1-10 mu m; the operating conditions of the microfiltration are the same as for the second dialysis. In the invention, the primary dialysate is stored in the industrial application process, and then secondary dialysis is carried out, so that in order to prevent impurities in a storage tank or a pipeline from clogging a membrane system, precise filtration is arranged before the secondary dialysis to filter out suspended particles possibly existing, and the safe and stable operation of a subsequent membrane process is ensured.
In the invention, the molecular weight cut-off of the nanofiltration membrane is preferably 450-550 Da; the pressure of the secondary dialysis is preferably 10-18 bar, and the temperature is preferably 48-55 ℃. In the invention, through secondary dialysis, the light transmittance of the feed liquid can reach more than 45%, the protein removal rate reaches 100%, the concentration multiple of the concentrated solution obtained through the secondary dialysis is 5-12, and the average flux can reach 15-20 LMH.
In the invention, the nanofiltration membrane is preferably a rolled nanofiltration membrane. The rolled nanofiltration membrane has the advantages of compact equipment structure, small occupied area, low energy consumption, low investment, high separation precision and the like.
In the present invention, after the second-stage dialysis is completed, preferably, the second-stage concentrated solution is further subjected to a second top washing, and the second top washing water is preferably a third-stage dialysate and/or an impurity solution filtered by a second reverse osmosis membrane (i.e., a dialysate obtained by filtering an impurity solution obtained by continuous chromatographic separation by the second reverse osmosis membrane); the volume of the second top washing water is preferably 10-20% of the first-level dialysate. In the present invention, it is preferable that the top washing solution obtained by the second top washing is dialyzed, and the obtained dialysate is combined with the dialysate obtained by the second-stage dialysis to be used as a second-stage dialysate, and the subsequent steps are performed; the dialysis conditions of the top wash solution were the same as the secondary dialysis. In the invention, the recovery rate of isoleucine can be improved by washing the secondary concentrated solution; the washed secondary concentrated solution can be discharged after environmental protection treatment.
After the secondary dialysate is obtained, the invention carries out continuous chromatographic separation on the secondary dialysate to obtain product liquid and impurity liquid. In the invention, the continuous chromatographic separation can separate isoleucine and salt to obtain a product liquid with low impurity content, and the obtained impurity liquid can be reused as top washing water of the primary concentrated solution and the secondary concentrated solution after being filtered by the second reverse osmosis membrane.
In the invention, the rejection rate of the second reverse osmosis membrane to sodium chloride is preferably more than or equal to 96 percent; the second reverse osmosis membrane is preferably made of polyamide; the pressure of the filtration is 16-18 bar, and the temperature is 30-50 ℃. In the invention, the water recovery rate of the impurity liquid can reach more than 82%, and the average flux of filtration can reach 25-30 LMH.
In the invention, the continuous chromatographic separation system preferably comprises an elution zone, an extension separation zone, a main separation zone and a post separation zone, each zone independently preferably comprises 2-5 resin columns, and the resin columns are preferably connected in series or in parallel; in the embodiment of the invention, the resin columns are connected in series.
In the embodiment of the present invention, the continuous chromatographic separation process is as shown in fig. 2, and includes an elution region, an extended separation region, a main separation region and a post-separation region, wherein the elution region includes 3 resin columns (i.e., resin columns No. 1, 4 and 7), the extended separation region includes 2 resin columns (i.e., resin columns No. 10 and 13), the main separation region includes 3 resin columns (i.e., resin columns No. 16, 19 and 22), the post-separation region includes 2 resin columns (i.e., resin columns No. 25 and 28), the resin columns of the functional regions are connected in series, each functional region is provided with a corresponding material pump to pump the material into the corresponding functional region, the material discharged from each functional region enters a transfer tank, and the four functional regions form an annular circulation system and are in a state of continuous rotation circulation; in the operation process, feed liquid is pumped into the main separation area by a No. 3 pump, isoleucine and impurities have different advancing speeds in resin, the advancing speed of isoleucine is slower, the advancing speed of impurities is faster, liquid containing the impurities enters the impurity cache tank, the isoleucine is gradually enriched on a resin column, the feed liquid is gradually consumed, a new material is pumped into the feed tank by a No. 5 pump, and the liquid level of the feed tank is kept stable; the No. 4 pump pumps the impurity-containing liquid stored in the impurity cache tank into the rear separation area, the movement speed of impurities such as pigment, salt and the like in the resin column is slower than that of water, the water can firstly enter the reuse water collection tank, the impurities such as pigment, salt and the like are gradually enriched on the resin column and finally still return to the impurity cache tank, and the No. 7 pump can pump a part of impurity-containing liquid in the impurity cache tank into the impurity collection tank to be impurity liquid; pumping isoleucine product buffer solution into the extended separation area by a No. 2 pump, enabling part of resin columns of the main separation area to step into the extended separation area, enabling part of buffer solution in the product buffer tank to enter the resin columns so as to further enrich isoleucine in the resin columns, enabling a small part of impurities which are not completely separated in the main separation area to enter the feeding buffer tank, and pumping part of products in the product buffer tank into a product collection tank by a No. 6 pump so as to obtain product solution; pumping pure water into an elution area by a No. 1 pump, enabling resin columns in the main separation area and the extension separation area to step into the area, enabling most of isoleucine products in the resin columns in the area to be isoleucine products, eluting the isoleucine products by the pure water at a certain flow rate, and enabling the eluted resin columns to circularly enter a rear separation area.
After the product liquid is obtained, the product liquid is subjected to three-stage dialysis through the first reverse osmosis membrane to obtain a three-stage concentrated liquid and a three-stage dialysate. In the invention, the third-stage dialysis is used for concentrating the product liquid to obtain a third-stage concentrated liquid with higher isoleucine concentration; the first reverse osmosis membrane is adopted for concentration, so that the advantage of low energy consumption is achieved; the obtained third-stage dialyzate can be directly recycled as top washing water of the first-stage concentrated solution and the second-stage concentrated solution.
In the invention, the rejection rate of the first reverse osmosis membrane to sodium chloride is preferably more than or equal to 96 percent; the first reverse osmosis membrane is preferably made of polyamide; the pressure of the three-stage dialysis is 12-16 bar, and the temperature is 40-52 ℃. In the invention, the concentration of isoleucine in the obtained tertiary concentrated solution can reach about 40g/L, and the average flux can reach 20-30 LMH. In the invention, the concentration multiple of the obtained three-stage concentrated solution is preferably 3-7 times.
In the present invention, the first reverse osmosis membrane is preferably a rolled reverse osmosis membrane. In the invention, the roll-type reverse osmosis membrane has the advantages of compact equipment structure, small occupied area, low energy consumption, small investment, high separation precision and the like.
According to the invention, the product liquid is preferably subjected to precise filtration before the third-stage dialysis; the aperture of the filter hole for the precision filtration is preferably 1-10 mu m; the operating conditions of the microfiltration process are the same as for the third-stage dialysis. In the invention, similar to the secondary dialysis precision filtration, in industry, before the tertiary dialysis, the feed liquid is stored, in order to avoid impurities existing in a storage tank and a pipeline from blocking a membrane system, the precision filtration is arranged to filter out suspended particles possibly existing, and the safe and stable operation of the subsequent membrane process is ensured.
After the third-level concentrated solution is obtained, the third-level concentrated solution is sequentially subjected to activated carbon decoloration, freeze crystallization and drying to obtain isoleucine.
In the invention, the specific steps of the activated carbon decolorization are as follows: mixing the third-stage concentrated solution with activated carbon, stirring, and performing suction filtration to obtain a decolorized third-stage concentrated solution; the feed-liquid ratio of the third-stage concentrated solution to the activated carbon is 3 g/L; the stirring time is preferably 20-30 min, the stirring speed is preferably 10-20 r/min, and the stirring can be stopped when the chroma is less than 5.
The freezing crystallization is not particularly limited in the invention, and a conventional freezing crystallization mode can be adopted.
The drying method is not particularly limited, and isoleucine with constant weight can be obtained.
In the present invention, the mode of the primary dialysis, the secondary dialysis and the tertiary dialysis is preferably cross-flow dialysis. In the invention, the adoption of cross-flow dialysis is an efficient, economic and reliable filtration mode.
The following examples are provided to illustrate the purification process of isoleucine fermentation broth of the present invention, but they should not be construed as limiting the scope of the present invention.
Example 1
The isoleucine hair liquid treated in this example is earthy yellow, pH is 6.6, conductivity is 22000 mus/cm, isoleucine content is 22g/L, and treatment amount is 100L.
Filtering isoleucine fermentation liquid with a filter with pore size of 50 μm, and performing primary dialysis at 50 deg.C under 4bar by using ceramic membrane with molecular weight cutoff of 50 KD; the concentration multiple of the concentrated solution obtained by the first-stage dialysis is 5.6 times; the light transmittance of the dialysate obtained by the primary dialysis is 12%;
carrying out first top washing on the concentrated solution obtained by the first-stage dialysis; the volume of water used for the first top washing is 40 percent of that of the isoleucine fermentation broth; mixing the dialysate obtained after the first top washing with dialysate obtained by the first-stage dialysis to obtain first-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the first top washing as first-stage concentrated solution;
performing precise filtration on the primary dialysate through a filter with the filter pore diameter of 10 μm, and performing secondary dialysis through a roll-type nanofiltration membrane with the molecular weight cutoff of 500Da, wherein the temperature of the secondary dialysis is 50 ℃, and the pressure is 18 bar; the concentration multiple of the concentrated solution obtained by the secondary dialysis is 10 times; the light transmittance of the dialysate obtained by the secondary dialysis is 40%;
carrying out second top washing on the concentrated solution obtained by the second-stage dialysis; the volume of water used for the second top washing is 20 percent of that of the first-stage liquid; mixing the dialysate obtained after the second top washing with dialysate obtained by the second-stage dialysis to obtain a second-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the second top washing as a second-stage concentrated solution;
carrying out continuous chromatographic separation on the secondary dialysate, wherein the flow of the continuous chromatographic separation is shown in figure 2, and obtaining a product liquid and an impurity liquid; the concentration of isoleucine in the obtained product liquid is 8g/L, and the light transmittance is 70%;
filtering the impurity liquid through a second roll type reverse osmosis membrane to be used as first top washing water and second top washing water; the filtering temperature is 50 ℃, the pressure is 18bar, the second roll type reverse osmosis membrane is made of polyamide, and the rejection rate of sodium chloride is more than or equal to 96%.
Performing precise filtration on the obtained product liquid through a filter with the filter pore diameter of 10 mu m, and performing three-stage dialysis through a first roll type reverse osmosis membrane, wherein the first roll type reverse osmosis membrane is made of polyamide, the rejection rate of sodium chloride is more than or equal to 96%, the temperature of the three-stage dialysis is 50 ℃, and the pressure is 18 bar; obtaining a third-stage dialysate and a third-stage concentrated solution after the third-stage dialysis; the tertiary dialysate can be used as first top washing water and second top washing water, and the light transmittance of the tertiary dialysate is 100%; the concentration multiple of the third-stage concentrated solution is 5 times, and the concentration of isoleucine in the third-stage concentrated solution is 30 g/L;
and (3) carrying out activated carbon decoloration on the third-stage concentrated solution, wherein the activated carbon decoloration comprises the following specific steps: adding activated carbon into the third-stage concentrated solution (the material-liquid ratio of the activated carbon to the third-stage concentrated solution is 3g/L), continuously stirring uniformly by using a stirrer, and performing suction filtration to obtain a decolorized third-stage concentrated solution when the chroma is less than 5;
and (4) carrying out freeze crystallization on the decolored third-level concentrated solution, and drying a solid obtained by freeze crystallization to obtain 2kg of isoleucine.
The calculated recovery rate of isoleucine is 90%, and the purity of the obtained isoleucine is 99.7% by adopting liquid chromatography detection.
Example 2
The isoleucine fermentation broth treated in this example was earthy yellow, had a pH of 6.5, an electrical conductivity of 25000. mu.s/cm, an isoleucine content of 21g/L, and a treatment capacity of 1000L.
Filtering isoleucine fermentation liquid with a filter with pore size of 50 μm, and performing primary dialysis at 55 deg.C under 4bar by ceramic membrane with molecular weight cutoff of 50 KD; the concentration multiple of the concentrated solution obtained by the first-stage dialysis is 6 times; the light transmittance of the dialysate obtained by the primary dialysis is 12%;
carrying out first top washing on the concentrated solution obtained by the first-stage dialysis; the volume of water used for the first top washing is 38 percent of that of the isoleucine fermentation broth; mixing the dialysate obtained after the first top washing with dialysate obtained by the first-stage dialysis to obtain first-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the first top washing as first-stage concentrated solution;
performing precise filtration on the primary dialysate through a filter with the filter pore diameter of 10 μm, and performing secondary dialysis through a roll-type nanofiltration membrane with the molecular weight cutoff of 500Da, wherein the temperature of the secondary dialysis is 52 ℃, and the pressure is 17 bar; the concentration multiple of the concentrated solution obtained by the secondary dialysis is 11 times; the light transmittance of the dialysate obtained by the secondary dialysis is 38%;
carrying out second top washing on the concentrated solution obtained by the second-stage dialysis; the volume of water used for the second top washing is 18 percent of that of the first-stage liquid; mixing the dialysate obtained after the second top washing with dialysate obtained by the second-stage dialysis to obtain a second-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the second top washing as a second-stage concentrated solution;
carrying out continuous chromatographic separation on the secondary dialysate, wherein the flow of the continuous chromatographic separation is shown in figure 2, and obtaining a product liquid and an impurity liquid; the concentration of isoleucine in the obtained product liquid is 8.2g/L, and the light transmittance is 69%;
filtering the impurity liquid through a second roll type reverse osmosis membrane to be used as first top washing water and second top washing water; the filtering temperature is 52 ℃, the pressure is 16bar, the second roll type reverse osmosis membrane is made of polyamide, and the rejection rate of sodium chloride is more than or equal to 96%.
Performing precise filtration on the obtained product liquid through a filter with the filter pore diameter of 10 mu m, and performing three-stage dialysis through a first roll type reverse osmosis membrane, wherein the first roll type reverse osmosis membrane is made of polyamide, the rejection rate of sodium chloride is more than or equal to 96%, the temperature of the three-stage dialysis is 52 ℃, and the pressure is 18 bar; obtaining a third-stage dialysate and a third-stage concentrated solution after the third-stage dialysis; the tertiary dialysate can be used as first top washing water and second top washing water, and the light transmittance of the tertiary dialysate is 100%; the concentration multiple of the third-stage concentrated solution is 5.5 times, and the concentration of isoleucine in the third-stage concentrated solution is 33 g/L;
and (3) carrying out activated carbon decoloration on the third-stage concentrated solution, wherein the activated carbon decoloration comprises the following specific steps: adding activated carbon into the third-stage concentrated solution (the material-liquid ratio of the activated carbon to the third-stage concentrated solution is 3g/L), continuously stirring uniformly by using a stirrer, and performing suction filtration to obtain a decolorized third-stage concentrated solution when the chroma is less than 5;
and (4) carrying out freeze crystallization on the decolored third-stage concentrated solution, and drying a solid obtained by freeze crystallization to obtain 18.48kg of isoleucine.
The calculated recovery rate of isoleucine is 88%, and the purity of the obtained isoleucine is 99.5% by adopting liquid chromatography detection.
Example 3
The isoleucine fermentation broth treated in this example was earthy yellow, had a pH of 6.9, an electrical conductivity of 24000. mu.s/cm, an isoleucine content of 23g/L, and a treatment capacity of 1000L.
Filtering isoleucine fermentation liquid with a filter with pore size of 50 μm, and performing primary dialysis at 60 deg.C under 3.6bar by ceramic membrane with molecular weight cutoff of 50 KD; the concentration multiple of the concentrated solution obtained by the first-stage dialysis is 6 times; the light transmittance of the dialysate obtained by the primary dialysis is 11%;
carrying out first top washing on the concentrated solution obtained by the first-stage dialysis; the volume of water used for the first top washing is 35 percent of that of the isoleucine fermentation broth; mixing the dialysate obtained after the first top washing with dialysate obtained by the first-stage dialysis to obtain first-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the first top washing as first-stage concentrated solution;
performing precise filtration on the primary dialysate through a filter with the filter pore diameter of 10 μm, and performing secondary dialysis through a roll-type nanofiltration membrane with the molecular weight cutoff of 500Da, wherein the temperature of the secondary dialysis is 55 ℃, and the pressure is 16 bar; the concentration multiple of the concentrated solution obtained by the secondary dialysis is 14 times; the light transmittance of the dialysate obtained by the secondary dialysis is 37%;
carrying out second top washing on the concentrated solution obtained by the second-stage dialysis; the volume of water used for the second top washing is 15 percent of that of the first-stage liquid; mixing the dialysate obtained after the second top washing with dialysate obtained by the second-stage dialysis to obtain a second-stage dialysate, and performing environment-friendly treatment on the concentrated solution obtained after the second top washing as a second-stage concentrated solution;
carrying out continuous chromatographic separation on the secondary dialysate, wherein the flow of the continuous chromatographic separation is shown in figure 2, and obtaining a product liquid and an impurity liquid; the concentration of isoleucine in the obtained product liquid is 8.8g/l, and the light transmittance is 66%;
filtering the impurity liquid through a second roll type reverse osmosis membrane to be used as first top washing water and second top washing water; the filtering temperature is 55 ℃, the pressure is 15bar, the second roll type reverse osmosis membrane is made of polyamide, and the rejection rate of sodium chloride is more than or equal to 96%.
Performing precise filtration on the obtained product liquid through a filter with the filter pore diameter of 10 mu m, and performing three-stage dialysis through a first roll type reverse osmosis membrane, wherein the first roll type reverse osmosis membrane is made of polyamide, the rejection rate of sodium chloride is not less than 96%, the temperature of the three-stage dialysis is 55 ℃, and the pressure is 15 bar; obtaining a third-stage dialysate and a third-stage concentrated solution after the third-stage dialysis; the tertiary dialysate can be used as first top washing water and second top washing water, and the light transmittance of the tertiary dialysate is 100%; the concentration multiple of the third-stage concentrated solution is 5 times, and the concentration of isoleucine in the third-stage concentrated solution is 35 g/L;
and (3) carrying out activated carbon decoloration on the third-stage concentrated solution, wherein the activated carbon decoloration comprises the following specific steps: adding activated carbon into the third-stage concentrated solution (the material-liquid ratio of the activated carbon to the third-stage concentrated solution is 3g/L), continuously stirring uniformly by using a stirrer, and performing suction filtration to obtain a decolorized third-stage concentrated solution when the chroma is less than 5;
and (4) carrying out freeze crystallization on the decolored third-stage concentrated solution, and drying a solid obtained by freeze crystallization to obtain 20.93kg of isoleucine.
The calculated recovery rate of isoleucine is 91%, and the purity of the obtained isoleucine is 99.6% by adopting a liquid chromatography method for detection.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (5)
1. The process for purifying the isoleucine fermentation liquor is characterized by comprising the following steps of:
(1) performing primary dialysis on the isoleucine fermentation broth through a ceramic membrane to obtain primary dialysate and primary concentrated solution;
(2) performing secondary dialysis on the primary dialysate through a nanofiltration membrane to obtain secondary dialysate and secondary concentrated solution;
(3) carrying out continuous chromatographic separation on the secondary dialysate to obtain a product liquid and an impurity liquid;
(4) performing third-stage dialysis on the product liquid through a first reverse osmosis membrane to obtain a third-stage concentrated liquid and a third-stage dialysate;
(5) sequentially decoloring the third-stage concentrated solution by using activated carbon, freezing, crystallizing and drying to obtain isoleucine;
performing precise filtration independently before the first-stage dialysis, the second-stage dialysis and the third-stage dialysis, and taking the filtrate for performing the first-stage dialysis, the second-stage dialysis and the third-stage dialysis, wherein the aperture of a filter hole of the precise filtration before the first-stage dialysis is 1-50 mu m, and the aperture of a filter hole of the precise filtration before the second-stage dialysis and the third-stage dialysis is 1-10 mu m independently;
after the first-stage dialysis is finished, carrying out first top washing on the first-stage concentrated solution, wherein water for the first top washing is a third-stage dialysate and/or an impurity solution filtered by a second reverse osmosis membrane; the volume of the first top washing water is 35-45% of that of the isoleucine fermentation broth;
after the second-stage dialysis is finished, second top washing is carried out on the second-stage concentrated solution, and water for the second top washing is third-stage dialysate and/or impurity liquid filtered by a second reverse osmosis membrane; the volume of the second top washing water is 10-20% of the first-level dialysate;
the continuous chromatographic separation system comprises an elution area, an extension separation area, a main separation area and a rear separation area, wherein the elution area comprises 3 resin columns, the extension separation area comprises 2 resin columns, the main separation area comprises 3 resin columns, the rear separation area comprises 2 resin columns, the resin columns of the functional areas are connected in series, the functional areas are provided with corresponding material pumps for pumping materials into the corresponding functional areas, the materials discharged from the functional areas enter a transfer tank, and the four functional areas form an annular circulation system and are in a state of continuous rotary circulation; in the operation process, feed liquid is pumped into the main separation area by a pump, liquid containing impurities enters the impurity cache tank, isoleucine is gradually enriched on the resin column, the feed liquid is gradually consumed, and a new material is pumped into the feed tank by the pump to maintain the liquid level of the feed tank to be stable; pumping the impurity-containing liquid stored in the impurity cache tank into a rear separation area, wherein water can firstly enter a reuse water collection tank, pigment and salt impurities are gradually enriched on a resin column and finally return to the impurity cache tank, and pumping a part of impurity-containing liquid in the impurity cache tank into the impurity collection tank by a pump to obtain impurity liquid; pumping the isoleucine product buffer solution into an extension separation area, enabling part of resin columns of a main separation area to step into the separation area, enabling part of buffer solution in a product buffer tank to enter the resin columns so as to further enrich isoleucine in the resin columns, enabling a small part of impurities which are not completely separated in the main separation area to enter a feeding buffer tank, and pumping part of products in the product buffer tank into a product collection tank by a pump to obtain product solution; pure water is pumped into the elution area, the resin columns in the main separation area and the extension separation area step into the area, most of the resin columns in the area are isoleucine products, the isoleucine products can be obtained by eluting with the pure water at a certain flow rate, and the eluted resin columns circularly enter the rear separation area.
2. The process for purifying an isoleucine fermentation broth according to claim 1, wherein the molecular weight cut-off of said ceramic membrane is 45-55 KD; the pressure of the primary dialysis is 3-4 bar, and the temperature is 40-60 ℃.
3. The purification process of isoleucine fermentation broth according to claim 1, wherein the molecular weight cut-off of the nanofiltration membrane is 450-550 Da; the pressure of the secondary dialysis is 10-18 bar, and the temperature is 48-55 ℃.
4. The process for purifying isoleucine fermentation broth of claim 1, wherein the rejection rate of said first reverse osmosis membrane to sodium chloride is not less than 96%; the pressure of the three-stage dialysis is 12-16 bar, and the temperature is 40-52 ℃.
5. The process for purifying isoleucine fermentation broth of claim 1, wherein the primary dialysis, secondary dialysis and tertiary dialysis are cross-flow dialysis.
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