CN116253657A - Method for extracting L-tyrosine from fermentation liquor - Google Patents

Method for extracting L-tyrosine from fermentation liquor Download PDF

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CN116253657A
CN116253657A CN202310301024.XA CN202310301024A CN116253657A CN 116253657 A CN116253657 A CN 116253657A CN 202310301024 A CN202310301024 A CN 202310301024A CN 116253657 A CN116253657 A CN 116253657A
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徐庆阳
王金多
陈志超
赵春光
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Tianjin University of Science and Technology
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Abstract

The utility model provides a method for extracting L-tyrosine from fermentation liquor, which comprises the steps of obtaining L-tyrosine crystals containing partial thalli and bacterial suspension containing saturated L-tyrosine solution through low-speed centrifugal separation; separating the bacterial suspension by high-speed centrifugation to obtain thalli and supernatant, wherein the supernatant is used as L-tyrosine crystals containing a small amount of thalli, which are obtained by mixed heavy suspension low-speed centrifugation, and performing low-speed centrifugation again to separate the crystals to obtain L-tyrosine crystals containing a small amount of thalli; slowly dissolving all L-tyrosine crystals in the obtained L-tyrosine crystal suspension by using acid liquor to obtain L-tyrosine solution containing a small amount of thalli, and carrying out high-speed centrifugal separation on supernatant and thalli, wherein thalli are recovered, and after the supernatant is decolorized by using active carbon, filtering the supernatant by using a plate frame to remove waste active carbon and filtrate; slowly adding ammonia water into the filtrate for continuous crystallization, performing high-speed centrifugal separation to obtain wet crystals, and drying to obtain high-yield and high-purity L-tyrosine crystals.

Description

Method for extracting L-tyrosine from fermentation liquor
Technical Field
The utility model relates to an extraction method of amino acid, in particular to a method for extracting L-tyrosine from fermentation liquor.
Background
L-tyrosine is one of the three major aromatic amino acids, an important conditionally essential amino acid. Has important functions in metabolism, growth and development of human and animals, and has been widely applied in the industries of medicine, food, feed, chemical industry, agriculture, cosmetics and the like.
The separation of products in the L-tyrosine industry not only requires that the final products meet the related requirements, but also requires that the loss of L-tyrosine is reduced, the purity of the products is improved, the water and electricity resources are saved, and the extraction cost is reduced in the separation process.
The national formulary requires that the purity of L-tyrosine is more than 99.0%, white crystals or crystalline powder is odorless, the specific rotation is-11.3 DEG to-12.1 DEG, and the light transmittance is not less than 95.0%. The utility model patent (N209338523U) provides a device for extracting L-tyrosine from fermentation liquor by a centrifugal method, but a large amount of fermentation liquor, pigment and thalli are still entrained in the crude L-tyrosine extracted according to the utility model, and the purity is to be improved.
Disclosure of Invention
The utility model aims to provide a method for extracting L-tyrosine from fermentation liquor.
In order to solve the technical problems, the technical scheme of the utility model is as follows:
a method for extracting L-tyrosine from fermentation broth comprises centrifuging at low speed to obtain bacterial suspension containing partial bacterial L-tyrosine crystals and saturated L-tyrosine solution; separating the bacterial suspension by high-speed centrifugation to obtain bacterial protein and supernatant, wherein the supernatant is used as L-tyrosine crystals containing a small amount of bacterial cells, which are obtained by mixed heavy suspension low-speed centrifugation, and the heavy suspension is centrifuged again at a low speed to obtain L-tyrosine crystals containing a small amount of bacterial cells; slowly dissolving all L-tyrosine crystals in the obtained L-tyrosine crystal suspension by using acid liquor to obtain an L-tyrosine solution containing a small amount of thalli, and carrying out high-speed centrifugal separation on supernatant and thalli, wherein thalli are recovered, and the supernatant is decolorized by using activated carbon and then filtered by a plate frame to remove waste activated carbon and filtrate; slowly adding ammonia water into the filtrate for continuous crystallization, performing high-speed centrifugal separation to obtain wet L-tyrosine crystals, and drying to obtain the L-tyrosine crystals.
Preferably, the method for extracting L-tyrosine from fermentation broth comprises the following specific steps:
(1) Centrifuging the fermentation liquor at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid A and a crystal A; the upper layer bacterial liquid A consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal A consists of bacterial proteins and L-tyrosine crystals;
(2) Carrying out high-speed centrifugation on the upper bacterial liquid A obtained in the step (1) at a centrifugal speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(3) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (2) with the crystal A to obtain a suspension A;
(4) Centrifuging the suspension A obtained in the step (3) at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid B and a crystal B; the upper layer bacterial liquid B consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal B is an L-tyrosine crystal still containing a small amount of bacterial cells;
(5) Centrifuging the upper layer bacterial liquid B obtained in the step (4) at a high speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(6) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (5) with the crystal B to obtain a suspension B;
(7) Centrifuging the suspension B in the step (6) at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid C and a crystal C; the upper bacterial liquid C consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal C is an L-tyrosine crystal still containing a small amount of bacterial cells;
(8) Centrifuging the upper layer bacterial liquid C obtained in the step (7) at a high speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(9) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (8) with the crystal C to obtain a suspension C;
(11) Slowly adding acid liquor into the suspension C in the step (10) to completely dissolve L-tyrosine crystals, thereby obtaining high-concentration L-tyrosine solution;
(12) Centrifuging the high-concentration L-tyrosine solution obtained in the step (11) at a high speed to obtain a supernatant A and a small amount of mycoprotein, and directly recovering the mycoprotein;
(13) Decolorizing the supernatant A obtained in the step (12) by using active carbon;
(14) Performing plate and frame filtration on the supernatant A mixed with the activated carbon obtained in the step (13) to obtain filtrate and waste activated carbon;
(15) Adding ammonia water into the filtrate flow obtained in the step (14) to adjust the pH value for continuous crystallization;
(16) Performing high-speed centrifugation after continuous crystallization in the step (15), and separating to obtain wet L-tyrosine crystals and mother liquor;
(17) Drying the wet L-tyrosine crystals obtained in the step (16) to obtain L-tyrosine crystals.
Preferably, in the method for extracting L-tyrosine from fermentation broth, the low-speed centrifugal rotation speed in the steps (1), (4) and (7) is 3000r/min.
Preferably, in the method for extracting L-tyrosine from fermentation broth, the high-speed centrifugal rotation speed in the steps (2), (5), (8) and (16) is 5000r/min.
Preferably, in the method for extracting L-tyrosine from fermentation broth, the acid liquor in the step (11) is dilute sulfuric acid, and the pH after adding the dilute sulfuric acid is controlled to be 2-3.
Preferably, in the method for extracting L-tyrosine from fermentation broth, the decolorizing temperature of the activated carbon in the step (13) is room temperature, and the stirring speed of the decolorizing tank is 200r/min.
Preferably, in the method for extracting L-tyrosine from fermentation broth, the ammonia water in the step (15) is 20% ammonia water, and the pH is controlled to be 5.6-5.7.
Preferably, the method for extracting L-tyrosine from fermentation broth as described above, wherein the mycoprotein is recovered in the step (12) for use as a feed protein.
Preferably, the method for extracting L-tyrosine from fermentation broth as described above, wherein the mother liquor in the step (16) is used for the preparation of the post-L-tyrosine fermentation medium.
Preferably, in the above method for extracting L-tyrosine from fermentation broth, the temperature of the drying process in the step (17) is 80 ℃.
The beneficial effects are that:
the method for extracting the L-tyrosine from the fermentation broth has the advantages of less loss of the L-tyrosine in the extraction process, high final yield and high purity of the obtained product; the extraction process does not use the traditional membrane separation and exchange process, and no waste liquid is discharged; the supernatant is subjected to crystal resuspension after centrifugation, fresh water is not used, and the water-saving effect is obvious; the mycoprotein produced by fermentation is recovered and used as feed, so that the emission of waste residue is reduced; the mother liquor obtained finally is rich in ammonium sulfate and is used as a fermentation medium or an organic fertilizer.
Drawings
FIG. 1 is a schematic diagram of L-tyrosine extraction scheme.
Detailed Description
In order to enable those skilled in the art to better understand the technical scheme of the present utility model, the technical scheme of the present utility model will be further described in detail below with reference to the specific embodiments.
The fermentation broth used in the following examples was obtained by fermentation using the strain and method described in the utility model patent of L-tyrosine genetic engineering bacterium and the method for producing L-tyrosine (application No. 201810963798.8).
Example 1
A method for extracting L-tyrosine from fermentation broth comprises the following specific steps:
a. taking 4L of L-tyrosine fermentation liquor (the L-tyrosine content is 45 g/L), and carrying out low-speed centrifugal separation on L-tyrosine crystals and most of thalli by using a centrifugal machine, wherein the centrifugal speed is 3000r/min, and the centrifugal time is 3min, so as to obtain crystal A and upper bacterial liquid A.
The upper layer bacterial liquid A obtained in b.a step does not contain L-tyrosine crystal, and is subjected to high-speed centrifugation at 5000r/min for 5min, and the bacterial cells and the solution containing saturated L-tyrosine are separated, and the bacterial cells are recovered for use as feed.
c. And b, stirring and suspending the crystal A obtained in the step a by using the solution containing the saturated L-tyrosine obtained in the step b to obtain a suspension A.
d. And (3) carrying out low-speed centrifugal separation on the L-tyrosine crystal and most of thalli by using a centrifugal machine on the suspension A, wherein the centrifugal speed is 3000r/min, and the centrifugal time is 3min, so as to obtain a crystal B and an upper bacterial liquid B.
The upper layer bacterial liquid B obtained in e.d step does not contain L-tyrosine crystal, and is subjected to high-speed centrifugation at 5000r/min for 10min, and the bacterial cells and the solution containing saturated L-tyrosine are separated, and the bacterial cells are recovered for use as feed.
f. And e, stirring and suspending the crystal B obtained in the step d by using the solution containing the saturated L-tyrosine obtained in the step e to obtain a suspension B.
g. Dilute sulfuric acid is added into the suspension B, and the pH is regulated to 2.0, so that L-tyrosine crystals are completely dissolved.
h. High-speed centrifugation was performed using the solution obtained in step g to obtain supernatant A.
i. The supernatant A was decolorized with activated carbon at room temperature with a stirring speed of 200r/min.
j. After the decoloring is finished, separating filtrate and waste active carbon by using a plate-frame filter.
k. Adding 20% ammonia water into the filtrate obtained in step j, adjusting the pH to 5.6-5.7, and carrying out continuous crystallization.
And I, obtaining the L-tyrosine wet crystal and mother liquor through high-speed centrifugal separation, wherein the centrifugal speed is 5000r/min, and the centrifugal time is 10min.
Drying wet L-tyrosine at 80 deg.C, and extracting with 92.3% yield and 99.1% purity.
Example 2
As shown in fig. 1, the difference compared to example 1 is that: and (3) carrying out low-speed centrifugation and thallus suspension repetition times for 3 times, obtaining suspension B, carrying out low-speed centrifugation to obtain upper layer bacterial liquid C and crystals C, carrying out high-speed centrifugation by using the upper layer bacterial liquid C to obtain thallus protein and a solution containing saturated L-tyrosine, stirring the suspension crystals C by using the solution containing saturated L-tyrosine, and continuing subsequent operations such as acid adding and dissolving. The suspension B is subjected to low-speed centrifugation, the centrifugation speed is 3000r/min, the centrifugation time is 3min, and the upper layer bacterial liquid C and the crystal C are obtained through separation; the upper bacterial liquid C consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal C is an L-tyrosine crystal still containing a small amount of bacterial cells; high-speed centrifugation is carried out on the obtained upper layer bacterial liquid C, the centrifugal speed is 5000r/min, the centrifugal time is 10min, and the bacterial protein and the solution containing saturated L-tyrosine are obtained through separation; and uniformly stirring the obtained solution containing saturated L-tyrosine and the crystal C to obtain a suspension C.
In this example, the extraction yield of L-tyrosine was 93.1% and the purity was 99.6% as compared with example 1.
Example 3
The difference from example 2 is that: and (3) carrying out low-speed centrifugation and thallus suspension repetition times of 4 times, obtaining suspension C, carrying out low-speed centrifugation to obtain upper layer bacterial liquid D and crystals D, carrying out high-speed centrifugation by using the upper layer bacterial liquid D to obtain thallus protein and a solution containing saturated L-tyrosine, stirring the suspension crystal D by using the solution containing saturated L-tyrosine, and continuing subsequent operations such as acid adding and dissolving. In this example, the recovery rate of L-tyrosine was 93.3% and the purity was 99.6% as compared with example 2.
Example 4
This embodiment differs from embodiment 2 in that: adding dilute sulfuric acid into the suspension C to dissolve L-tyrosine, and controlling the pH value to be 3.0. In this example, the extraction yield of L-tyrosine was 93.2% and the purity was 99.6% as compared with example 2.
Example 5
This embodiment differs from embodiment 2 in that: adding dilute sulfuric acid into the suspension C to dissolve L-tyrosine, and controlling the pH value to be 4.0. In this example, the extraction yield of L-tyrosine was 92.7% and the purity was 99.5% as compared with example 2.
The foregoing is merely a preferred embodiment of the present utility model, and it should be noted that, for those skilled in the art, it is possible to make several modifications and alterations without departing from the principles of the present utility model, and the steps of constructing the strain of the present utility model are not sequential, and those skilled in the art should consider the scope of the present utility model as modifications and alterations of the strain according to the method of the present utility model or based on the method.

Claims (10)

1. A method for extracting L-tyrosine from fermentation broth, which is characterized by comprising the following steps: firstly, carrying out low-speed centrifugal separation to obtain bacterial suspension containing partial bacterial L-tyrosine crystals and saturated L-tyrosine solution; separating the bacterial suspension by high-speed centrifugation to obtain bacterial protein and supernatant, wherein the supernatant is used as L-tyrosine crystals containing a small amount of bacterial cells, which are obtained by mixed heavy suspension low-speed centrifugation, and the heavy suspension is centrifuged again at a low speed to obtain L-tyrosine crystals containing a small amount of bacterial cells; slowly dissolving all L-tyrosine crystals in the obtained L-tyrosine crystal suspension by using acid liquor to obtain an L-tyrosine solution containing a small amount of thalli, and carrying out high-speed centrifugal separation on supernatant and thalli, wherein thalli are recovered, and the supernatant is decolorized by using activated carbon and then filtered by a plate frame to remove waste activated carbon and filtrate; slowly adding ammonia water into the filtrate for continuous crystallization, performing high-speed centrifugal separation to obtain wet L-tyrosine crystals, and drying to obtain the L-tyrosine crystals.
2. The method for extracting L-tyrosine from fermentation broth according to claim 1, wherein: the method comprises the following specific steps:
(1) Centrifuging the fermentation liquor at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid A and a crystal A; the upper layer bacterial liquid A consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal A consists of bacterial proteins and L-tyrosine crystals;
(2) Carrying out high-speed centrifugation on the upper bacterial liquid A obtained in the step (1) at a centrifugal speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(3) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (2) with the crystal A to obtain a suspension A;
(4) Centrifuging the suspension A obtained in the step (3) at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid B and a crystal B; the upper layer bacterial liquid B consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal B is an L-tyrosine crystal still containing a small amount of bacterial cells;
(5) Centrifuging the upper layer bacterial liquid B obtained in the step (4) at a high speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(6) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (5) with the crystal B to obtain a suspension B;
(7) Centrifuging the suspension B in the step (6) at a low speed, wherein the centrifugal speed is 2000-3000r/min; separating to obtain an upper bacterial liquid C and a crystal C; the upper bacterial liquid C consists of a solution containing saturated L-tyrosine and bacterial proteins, and the crystal C is an L-tyrosine crystal still containing a small amount of bacterial cells;
(8) Centrifuging the upper layer bacterial liquid C obtained in the step (7) at a high speed of 5000-6000r/min, and separating to obtain bacterial protein and a solution containing saturated L-tyrosine;
(9) Uniformly stirring the solution containing saturated L-tyrosine obtained in the step (8) with the crystal C to obtain a suspension C;
(11) Slowly adding acid liquor into the suspension C in the step (10) to completely dissolve L-tyrosine crystals, thereby obtaining high-concentration L-tyrosine solution;
(12) Centrifuging the high-concentration L-tyrosine solution obtained in the step (11) at a high speed to obtain a supernatant A and a small amount of mycoprotein, and directly recovering the mycoprotein;
(13) Decolorizing the supernatant A obtained in the step (12) by using active carbon;
(14) Performing plate and frame filtration on the supernatant A mixed with the activated carbon obtained in the step (13) to obtain filtrate and waste activated carbon;
(15) Adding ammonia water into the filtrate flow obtained in the step (14) to adjust the pH value for continuous crystallization;
(16) Performing high-speed centrifugation after continuous crystallization in the step (15), and separating to obtain wet L-tyrosine crystals and mother liquor;
(17) Drying the wet L-tyrosine crystals obtained in the step (16) to obtain L-tyrosine crystals.
3. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the low-speed centrifugal rotating speed in the steps (1), (4) and (7) is 3000r/min.
4. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the high-speed centrifugal rotating speed in the steps (2), (5), (8) and (16) is 5000r/min.
5. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: in the step (11), the acid liquor is dilute sulfuric acid, and the pH value after the dilute sulfuric acid is added is controlled to be 2-3.
6. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: and (3) the decolorizing temperature of the activated carbon in the step (13) is room temperature, and the stirring speed of the decolorizing tank is 200r/min.
7. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the ammonia water in the step (15) is 20% ammonia water, and the pH is controlled to be 5.6-5.7.
8. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the mycoprotein in step (12) is recovered for use as a feed protein.
9. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the mother liquor in the step (16) is used for preparing the post-L-tyrosine fermentation medium.
10. The method for extracting L-tyrosine from fermentation broth according to claim 2, wherein: the temperature of the drying process in the step (17) is 80 ℃.
CN202310301024.XA 2023-03-27 2023-03-27 Method for extracting L-tyrosine from fermentation liquor Pending CN116253657A (en)

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