CN111073917A - Circulating fermentation and separation extraction method for tyrosine - Google Patents
Circulating fermentation and separation extraction method for tyrosine Download PDFInfo
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- 238000000855 fermentation Methods 0.000 title claims abstract description 122
- 230000004151 fermentation Effects 0.000 title claims abstract description 122
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 title claims abstract description 65
- 238000000926 separation method Methods 0.000 title claims abstract description 51
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 title claims abstract description 49
- 238000000605 extraction Methods 0.000 title claims abstract description 37
- 239000007788 liquid Substances 0.000 claims abstract description 57
- 230000001580 bacterial effect Effects 0.000 claims abstract description 40
- 238000000034 method Methods 0.000 claims abstract description 40
- 239000013078 crystal Substances 0.000 claims abstract description 29
- 239000000919 ceramic Substances 0.000 claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 claims abstract description 24
- 238000000464 low-speed centrifugation Methods 0.000 claims abstract description 21
- 238000005374 membrane filtration Methods 0.000 claims abstract description 14
- 238000005119 centrifugation Methods 0.000 claims description 22
- 239000002253 acid Substances 0.000 claims description 7
- 238000011218 seed culture Methods 0.000 claims description 7
- 239000001963 growth medium Substances 0.000 claims description 5
- 241001052560 Thallis Species 0.000 abstract description 8
- 238000000703 high-speed centrifugation Methods 0.000 abstract 1
- 229960004441 tyrosine Drugs 0.000 description 43
- 239000012528 membrane Substances 0.000 description 14
- 230000008676 import Effects 0.000 description 12
- 241000894006 Bacteria Species 0.000 description 10
- 239000007791 liquid phase Substances 0.000 description 9
- 238000001914 filtration Methods 0.000 description 7
- 230000008569 process Effects 0.000 description 7
- 238000010586 diagram Methods 0.000 description 5
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 238000010353 genetic engineering Methods 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 238000005086 pumping Methods 0.000 description 3
- 238000004064 recycling Methods 0.000 description 3
- 239000007790 solid phase Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 239000002028 Biomass Substances 0.000 description 2
- 230000002411 adverse Effects 0.000 description 2
- 230000008878 coupling Effects 0.000 description 2
- 238000010168 coupling process Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 238000005265 energy consumption Methods 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000005526 G1 to G0 transition Effects 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 125000001477 organic nitrogen group Chemical group 0.000 description 1
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N phenol group Chemical group C1(=CC=CC=C1)O ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
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- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/04—Alpha- or beta- amino acids
- C12P13/22—Tryptophan; Tyrosine; Phenylalanine; 3,4-Dihydroxyphenylalanine
- C12P13/225—Tyrosine; 3,4-Dihydroxyphenylalanine
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/40—Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C227/00—Preparation of compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C227/38—Separation; Purification; Stabilisation; Use of additives
- C07C227/40—Separation; Purification
- C07C227/42—Crystallisation
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Abstract
The invention provides a circulating fermentation and separation extraction method of tyrosine, which comprises the steps of firstly fermenting to obtain tyrosine fermentation liquor, then separating most of tyrosine crystals and bacterial liquid through two-stage low-speed centrifugation, carrying out high-speed centrifugation or ceramic membrane filtration and concentration on the bacterial liquid, inoculating the concentrated thalli into another fermentation tank, and carrying out fermentation of a new batch. The method can primarily separate out most of tyrosine product crystals, and simultaneously, the thallus can be recycled, thereby improving the utilization rate of equipment and the production efficiency, compressing the non-production time and saving the energy resources.
Description
Technical Field
The invention relates to an extraction method of amino acid, in particular to a circulating fermentation and separation extraction method of tyrosine.
Background
L-tyrosine (L-tyrosine, L-Tyr) is a kind of aromatic α -amino-3-p-hydroxyphenylpropionic acid with phenolic hydroxyl group, is white crystalline powder, and is crystallized from water into needle-like or sheet-like body with relative density of 1.456(20 ℃), isoelectric point of 5.66 and water solubility of 0.45g/L (25 ℃), and has wide application in food, medicine, chemical industry and other fields.
The circulating fermentation can be a technology of inoculating a new fermentation culture medium to perform the next batch of fermentation under proper conditions by utilizing the thalli cultured in the last batch of fermentation culture. The application of the circulating fermentation technology can omit the seed culture process of the next batch of fermentation, shorten the time from the lag phase to the stabilization phase in the next batch of fermentation stage, save the time cost, save the energy resource, compress the production cost and improve the production efficiency.
Utilizing the patent: the fermentation experiment of the strain and the method in the L-tyrosine genetic engineering bacterium and the method for producing the L-tyrosine (application number CN201810963798.8) shows that the tyrosine fermentation process belongs to a non-growth coupling type, namely, the tyrosine starts to be synthesized in a large amount after the bacterium reaches a stabilizer, the culture time of the bacterium can be shortened by circulating fermentation, and the stable period can be reached quickly; meanwhile, because the solubility of tyrosine is very low, tyrosine is accumulated and crystallized in a large amount in fermentation liquor when being synthesized in a large amount, and the primary separation and extraction of a tyrosine product are completed by utilizing the density difference of crystals and bacterial liquid and performing the primary separation of the crystals and the bacterial liquid through a centrifugal technology; the thallus in the bacteria liquid can be concentrated by a centrifugal method or a membrane filtration method, and toxic and harmful substances such as hydrochloric acid and the like in part of the bacteria liquid are removed, so that healthy and safe thallus seeds are inoculated in the next batch of fermentation. Therefore, the circulating fermentation has great application potential for improving the production efficiency of tyrosine.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for circulating fermentation, separation and extraction of tyrosine, so as to realize the recycling of thalli.
In order to solve the technical problems, the technical scheme of the invention is as follows:
a circulating fermentation and separation extraction method of tyrosine comprises the following steps:
(1) performing seed culture and fermentation, and obtaining tyrosine fermentation liquor after the fermentation is finished, wherein the seed culture and fermentation method is performed by adopting an L-tyrosine genetic engineering bacterium in application number CN201810963798.8 and the strain and method in the method for producing L-tyrosine, and the fermentation is performed in a first fermentation tank;
(2) allowing the fermentation liquor obtained in the step (1) to enter a primary low-speed disc centrifuge from a first fermentation tank through a centrifugal pump for primary low-speed centrifugation, separating crystals and bacterial liquid, and using the disc centrifuge as a nozzle type;
(3) allowing the bacterial liquid obtained in the step (2) to enter a secondary disc type centrifuge through a centrifugal pump for secondary low-speed centrifugation, and further separating crystals in the bacterial liquid, wherein the centrifugation conditions are consistent with those in the step (2); because the amount of tyrosine crystals in the bacterial liquid after the secondary low-speed centrifugation is very small, the bacterial liquid is not further separated due to the cost;
(4) pumping the bacterial liquid obtained in the step (3) into a high-speed disc type centrifuge or a ceramic membrane filtering component for concentration, wherein the concentration method adopts a centrifugation method or a ceramic membrane filtering method; the purpose of concentration is as follows: harmful substances such as hydrochloric acid and the like remained in the bacterial liquid due to long-time fermentation culture are removed, and the influence of the components of the original culture medium on the next batch of fermentation culture medium is reduced;
(5) and (3) inoculating the thallus concentrated in the step (4) into a fermentation tank II containing a fermentation culture medium for the next batch of fermentation, wherein the fermentation method is the same as that in the step (1).
Preferably, in the above method for circulating fermentation and separation and extraction of tyrosine, the determination criterion of the end point of the fermentation in step (1) is that the acid production rate is 0.25-1g/L/h, if the acid production rate is higher than the criterion, the fermentation of the batch is insufficient, and the production efficiency is reduced, and if the acid production rate is lower than the criterion, the bacterial cells are aged, and the next round of fermentation is adversely affected.
Preferably, in the above method for circulating fermentation and separation and extraction of tyrosine, the centrifugation conditions in step (2) are: the separation factor Fr is 100-; if the separation factor or the centrifugation time is more than the range, excessive thalli mixed in the crystal are easily caused, and the yield of the thalli and the purity of the crystal are reduced; if the separation factor or the centrifugation time is less than the range, the separation of the crystal and the bacterial liquid is easy to be caused, and the extraction yield is reduced.
Preferably, in the above method for circulating fermentation and separation extraction of tyrosine, the crystal volume after the first-stage low-speed centrifugation in step (2): the volume ratio of the bacterial liquid is 1: 2-4, because the separation factor is too small, the bacteria liquid obtained by the first-stage low-speed centrifugation still contains a part of crystals, and the bacteria obtained by the first-stage low-speed centrifugation needs to be subjected to second-stage low-speed centrifugation for ensuring the extraction yield, and then the crystals are further collected.
Preferably, in the above method for circulating fermentation and separation extraction of tyrosine, after the second-stage low-speed centrifugation in step (3), the crystal volume is: the volume ratio of the bacterial liquid is 1: 9-20.
Preferably, in the above method for circulating fermentation and separation and extraction of tyrosine, the centrifugation method in step (4) uses a disk-type centrifuge, and the centrifugation conditions are as follows: separating for 1-3min with separation factor Fr being 1500-; centrifugal concentration ratio 1: 30-50.
According to the circulating fermentation and separation extraction method for tyrosine, the bacteria liquid is concentrated by the centrifugal method in the step (4), the solid-liquid separation effect is high, the concentration ratio is high, but the energy consumption of a centrifugal machine is easily overhigh due to the high separation factor, and the working noise is high; if the centrifugal condition is less than the standard, the yield of thalli is low, the inoculation amount of the next batch of fermentation is small, and the production efficiency is reduced; if the standard is larger than the standard, energy consumption is wasted, and the production cost is increased.
The equipment used in the method for circulating fermentation and separation extraction of tyrosine corresponding to the centrifugal method concentrated bacterial liquid comprises: the fermentation tank I, the low-speed disc centrifuge of one-level, the low-speed disc centrifuge of second grade, high-speed disc centrifuge, fermentation tank II and centrifugal pump and connecting tube, valve etc.. Wherein fermentation tank bottom links to each other through centrifugal pump and one-level low-speed disk centrifuge's feed liquid import, and one-level low-speed disk centrifuge's liquid phase export passes through the centrifugal pump and links to each other with second grade low-speed disk centrifuge feed liquid import, and second grade low-speed disk centrifuge's liquid phase export passes through the centrifugal pump and links to each other with high-speed disk centrifuge's feed liquid import, and high-speed disk centrifuge's solid phase export passes through the centrifugal pump and links to each other with No. two fermentation tank tops.
Preferably, in the above method for circulating fermentation and separation and extraction of tyrosine, the bacterial liquid is concentrated in the step (4) by ceramic membrane filtration, the ceramic membrane filtration condition is that the working pressure is 0.2-0.6MPa, and the concentration ratio is 1: 3-5, if the filtering condition of the ceramic membrane is more than the range, membrane blockage is easy to cause, membrane pollution is caused, and the water consumption for membrane cleaning is increased; if the concentration is less than the standard, the concentration cannot be achieved, and the next fermentation is adversely affected.
The equipment used in the method for circulating fermentation and separation extraction of tyrosine corresponding to the ceramic membrane filtration concentrated bacterial liquid comprises the following steps: the fermentation tank comprises a first-stage low-speed disc type centrifugal machine, a second-stage low-speed disc type centrifugal machine, a ceramic membrane filtering component, a second-stage fermentation tank, a centrifugal pump, a connecting pipeline, a valve and the like. Wherein: a fermentation cylinder bottom links to each other through centrifugal pump and one-level low-speed disc centrifuge's feed liquid import, and one-level low-speed disc centrifuge's liquid phase export passes through the centrifugal pump and links to each other with second grade low-speed disc centrifuge feed liquid import, and second grade low-speed disc centrifuge's liquid phase export passes through the centrifugal pump and links to each other with ceramic membrane filter element's feed liquid import, and ceramic membrane filter element's dense solution export passes through the centrifugal pump and links to each other with No. two fermentation cylinder tops.
The steps and the equipment are carried out under the condition of positive pressure in order to ensure the sterile environment, and the feed liquid is sterilized in advance before entering the next equipment.
Preferably, in the above tyrosine recycling fermentation and separation and extraction method, the fermentation medium in the step (5) is the tyrosine fermentation medium described in the patent (application No. CN 201810963798.8).
The invention has the beneficial effects that:
the circulating fermentation and separation extraction method of tyrosine has the following advantages:
(1) the utilization rate of the equipment is improved. The fermentation thalli is recycled, the seed culture time (generally 6-8h of the seed culture period) is saved, and the time from a lag phase to a stationary phase in a fermentation stage (from 12-14h to 5-7h) is shortened, so that the non-production time of a fermentation tank is greatly reduced, and the utilization rate of equipment is improved.
(2) And energy resources are saved. Large quantities of expensive organic nitrogen sources have to be put into operation before seed cultivation and fermentation to obtain large biomass in a short time, during which time the operation of various production and ancillary equipment also consumes large amounts of energy. The application of the circulating fermentation technology reduces the production cost.
(3) The automation degree of the equipment is high. The disc centrifuge or ceramic membrane filtering equipment is mature and stable equipment, can be sterilized, accords with GMP certification, is aseptically operated in the whole fermentation process, automatically monitors process parameters by a computer, and is favorable for fine management.
(4) The contamination probability is reduced. In fermentation production, in order to obtain sufficient biomass, the culture needs to be expanded step by step, and the strain is easy to infect in the process of transplanting.
(5) The production efficiency is improved. In the process of circulating fermentation, the process from the collection of the thalli from the batch of fermentation liquor to the inoculation of the next batch of fermentation liquor can be regarded as a seed transferring process. The preliminary separation and extraction of the tyrosine crystal product are realized while the seed is transferred, and the fermentation coupling extraction method improves the production efficiency.
Drawings
FIG. 1 is a schematic process flow diagram of the circulating fermentation and separation extraction method of tyrosine according to the invention.
FIG. 2 is a schematic flow diagram of the apparatus for centrifugal concentration according to the present invention.
FIG. 3 is a schematic flow diagram of an apparatus for concentration by ceramic membrane filtration according to the present invention.
FIG. 4 is a schematic diagram of a disk centrifuge of the apparatus for concentration by centrifugation according to the present invention.
FIG. 5 is a schematic structural diagram of a ceramic membrane filtration module corresponding to the ceramic membrane filtration concentration apparatus of the present invention.
In the figure: 1. the fermentation tank I, the first-stage low-speed disc type centrifuge, the second-stage low-speed disc type centrifuge, the high-speed disc type centrifuge, the fermentation tank II, the centrifugal pump 6 and the ceramic membrane filtering component 7
Detailed Description
The technical solution of the present invention is further described with reference to the following specific examples.
Example 1
A method for circulating fermentation and separation extraction of tyrosine, as shown in figure 1, figure 2 and figure 4, comprises the following steps:
(1) utilizing the patent: an L-tyrosine genetic engineering bacterium and a method for producing L-tyrosine (application number CN201810963798.8) thereof carry out seed culture and fermentation (tyrosine fermentation is carried out in a fermentation tank I), and after the fermentation is finished, tyrosine fermentation liquor is obtained;
(2) pumping the fermentation liquor obtained in the step (1) from the bottom of a first fermentation tank into a first-stage low-speed disc type centrifuge through a centrifugal pump to perform first-stage low-speed centrifugation, and separating crystals and bacterial liquid, wherein the centrifuge uses a disc type centrifuge;
(3) pumping the bacterial liquid obtained in the step (2) into a secondary low-speed disc centrifuge through a centrifugal pump for secondary low-speed centrifugation, further separating crystals in the bacterial liquid, wherein the centrifugation conditions are consistent with those in the step (2);
(4) concentrating the bacterial liquid obtained in the step (3), wherein a centrifugal method is adopted in the concentration method;
(5) inoculating the concentrated thallus obtained in the step (4) into another fermentation tank containing a fermentation medium (see the tyrosine fermentation medium in application No. CN201810963798.8) to perform the next batch of fermentation, wherein the fermentation method is the same as that of the step (1); wherein,
the acid production rate at the end point of the fermentation in the step (1) is 0.8 g/L/h;
adopting a disk-type centrifuge as a nozzle type in the steps (2) and (3), wherein the centrifugation conditions are as follows: separating factor Fr 150, centrifuging for 3 min; when the centrifugal machine works, the crystals are discharged from a solid-phase outlet of the centrifugal machine, and the bacterial liquid is discharged from a liquid-phase outlet of the centrifugal machine, so that the separation of the crystals and the bacterial liquid is realized;
the crystal volume after the first-stage low-speed centrifugation in the step (2): the volume ratio of the bacterial liquid is 1; 3;
after the second-stage low-speed centrifugation in the step (3), the crystal volume is as follows: the volume ratio of the bacterial liquid is 1: 20;
the centrifugation method in the step (4) uses a high-speed disc centrifuge, and the centrifugation conditions are as follows: separation factor Fr is 1600, separation time 3min, centrifugal concentration ratio 1: 40.
as shown in fig. 2 and 4, the device used in the tyrosine circulating fermentation and separation extraction method corresponding to the centrifugal method concentrated bacterial liquid in the step (4) comprises: the fermentation tank comprises a first-stage low-speed disc type centrifuge 1, a first-stage low-speed disc type centrifuge 2, a second-stage low-speed disc type centrifuge 3, a high-speed disc type centrifuge 4, a second-stage fermentation tank 5, a centrifugal pump 6, connecting pipelines, valves and the like. Wherein: a fermentation cylinder bottom links to each other through centrifugal pump and one-level low-speed disk centrifuge's feed liquid import, and one-level low-speed disk centrifuge's liquid phase export passes through the centrifugal pump and links to each other with second grade low-speed disk centrifuge feed liquid import, and second grade low-speed disk centrifuge's liquid phase export passes through the centrifugal pump and links to each other with high-speed disk centrifuge's feed liquid import, and high-speed disk centrifuge's solid phase export passes through the centrifugal pump and links to each other with No. two fermentation cylinder tops.
Example 2
A circulating fermentation and separation and extraction method of tyrosine, which is basically the same as the method of the embodiment 1, as shown in figure 1 and figure 3, and is characterized in that:
and (2) the acid production rate of the fermentation end point in the step (1) is 0.4 g/L/h.
A disc-type centrifuge is adopted in the step (2), and the centrifugation conditions are as follows: the separation factor Fr is 230, centrifugation time 2 min.
The crystal volume after the first-stage low-speed centrifugation in the step (2): the volume of the bacterial liquid is 1; 2.
after the second-stage low-speed centrifugation in the step (3), the crystal volume is as follows: the volume of the bacterial liquid is 1: 15.
the centrifugation method in the step (4) uses a disc-type centrifuge, and the centrifugation conditions are as follows: separation factor Fr ═ 2000, separation 1min, centrifugal concentration ratio 1: 50.
example 3
A circulating fermentation and separation extraction method of tyrosine, which is basically the same as the embodiment 1, except that:
and (2) the acid production rate of the fermentation end point in the step (1) is 0.7 g/L/h.
Adopting a disc centrifuge in the steps (2) and (3), wherein the centrifugation conditions are as follows: the separation factor Fr was 120 and the centrifugation time was 5 min.
The crystal volume after the first-stage low-speed centrifugation in the step (2): the volume of the bacterial liquid is 1; 2.5.
after the second-stage low-speed centrifugation in the step (3), the crystal volume is as follows: the volume of the bacterial liquid is 1: 12.
and (4) concentrating the bacterial liquid by adopting a ceramic membrane filtration method, wherein the ceramic membrane filtration condition is that the working pressure is 0.5MPa, and the concentration ratio is 1: 3.
as shown in fig. 3-5, the device used in the tyrosine circulating fermentation and separation extraction method corresponding to the ceramic membrane filtration method for concentrating the bacterial liquid in the step (4) comprises: the fermentation tank comprises a first fermentation tank 1, a first-stage low-speed disc type centrifuge 2, a second-stage low-speed disc type centrifuge 3, a ceramic membrane filtering component 7, a second fermentation tank 5, a centrifugal pump 6, connecting pipelines, valves and the like. Wherein: a fermentation cylinder bottom links to each other through centrifugal pump and one-level low-speed disc centrifuge's feed liquid import, and one-level low-speed disc centrifuge's liquid phase export links to each other through centrifugal pump and second grade low-speed disc centrifuge feed liquid import, and second grade low-speed disc centrifuge's liquid phase export passes through the feed liquid import that centrifugal pump and ceramic membrane filter assembly link to each other, and ceramic membrane filter assembly's dense solution export links to each other with No. two fermentation cylinder tops through the centrifugal pump.
The above detailed description of the tyrosine recycling fermentation and separation extraction process with reference to the examples is illustrative and not restrictive, and several examples are listed according to the limited scope, therefore, changes and modifications without departing from the general concept of the present invention shall fall within the protection scope of the present invention.
Claims (7)
1. A tyrosine circulating fermentation and separation extraction method is characterized in that:
(1) carrying out seed culture and fermentation, and obtaining tyrosine fermentation liquor after the fermentation is finished;
(2) performing primary low-speed centrifugation on the fermentation liquor obtained in the step (1), and separating crystals and bacterial liquid, wherein a disc-type centrifuge is used as the centrifuge;
(3) performing secondary low-speed centrifugation on the bacterial liquid obtained in the step (2), further separating crystals in the bacterial liquid, wherein the centrifugation conditions are consistent with those in the step (2);
(4) concentrating the bacterial liquid obtained in the step (3), wherein the concentration method adopts a centrifugal method or a ceramic membrane filtration method;
(5) and (3) inoculating the thallus concentrated in the step (4) into another fermentation tank containing a fermentation culture medium for the next batch of fermentation, wherein the fermentation method is the same as that in the step (1).
2. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: the judgment standard of the fermentation end point in the step (1) is that the acid production rate is 0.25-1 g/L/h.
3. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: the centrifugation conditions in the step (2) are as follows: the separation factor Fr is 100-.
4. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: the volume of the crystals after the first-stage low-speed centrifugation in the step (2) is as follows: the volume ratio of the bacterial liquid is 1: 2-4.
5. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: after the second-stage low-speed centrifugation in the step (3), the crystal volume is as follows: the volume ratio of the bacterial liquid is 1: 9-20.
6. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: the centrifugation method in the step (4) uses a disc-type centrifuge, and the centrifugation conditions are as follows: separating for 1-3min under the condition that the separation factor Fr is 1600-; centrifugal concentration ratio 1: 30-50.
7. The circulating fermentation and separation and extraction method of tyrosine according to claim 1, characterized in that: and (3) concentrating the bacterial liquid by a ceramic membrane filtration method in the step (4), wherein the ceramic membrane filtration condition is that the working pressure is 0.2-0.6MPa, and the concentration ratio is 1: 3-5.
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| CN116253657A (en) * | 2023-03-27 | 2023-06-13 | 天津科技大学 | Method for extracting L-tyrosine from fermentation liquor |
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