CA1215069A - Method of isolating l-trytophan - Google Patents
Method of isolating l-trytophanInfo
- Publication number
- CA1215069A CA1215069A CA000444165A CA444165A CA1215069A CA 1215069 A CA1215069 A CA 1215069A CA 000444165 A CA000444165 A CA 000444165A CA 444165 A CA444165 A CA 444165A CA 1215069 A CA1215069 A CA 1215069A
- Authority
- CA
- Canada
- Prior art keywords
- tryptophan
- exchange resin
- microorganism
- ion exchange
- produced
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12P13/227—Tryptophan
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D209/00—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D209/02—Heterocyclic compounds containing five-membered rings, condensed with other rings, with one nitrogen atom as the only ring hetero atom condensed with one carbocyclic ring
- C07D209/04—Indoles; Hydrogenated indoles
- C07D209/10—Indoles; Hydrogenated indoles with substituted hydrocarbon radicals attached to carbon atoms of the hetero ring
- C07D209/18—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
- C07D209/20—Radicals substituted by carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals substituted additionally by nitrogen atoms, e.g. tryptophane
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/02—Separating microorganisms from their culture media
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12P13/06—Alanine; Leucine; Isoleucine; Serine; Homoserine
Abstract
Title of the Invention METHOD OF ISOLATING L-TRYPTOPHAN
Abstract of the Disclosure When an L-tryptophan-containing reaction mixture produced by a reaction involving the use of a microorganism is treated with an H-type strongly acidic cation exchange resin without removing the microorganism from it, the L-tryptophan is adsorbed on the ion exchange resin and the microorganism is floc-culated on the ion exchange resin. The flocculated microorganism is easily removed by washing the ion exchange resin. The adsorbed L-tryptophan is eluted from the ion exchange resin and purified to a high purity.
Abstract of the Disclosure When an L-tryptophan-containing reaction mixture produced by a reaction involving the use of a microorganism is treated with an H-type strongly acidic cation exchange resin without removing the microorganism from it, the L-tryptophan is adsorbed on the ion exchange resin and the microorganism is floc-culated on the ion exchange resin. The flocculated microorganism is easily removed by washing the ion exchange resin. The adsorbed L-tryptophan is eluted from the ion exchange resin and purified to a high purity.
Description
~L2~5~
This invention relates to a method of effuse-entry isolating L-tryptophan from a reaction mixture obtained during the production of L-tryptophan my utilizing a microorganism.
Many methods have been reported previously for isolating an L-amino acid by an ion exchange resin from an L-amino acid-containing reaction mixture produced by utilizing a microorganism. For example, there have been known the separation of basic amino acids by carboxylic acid-type cation exchange resins (U. So Patent No. 2,549,378), the separation of acidic amino acids by weakly basic anion exchange resins (D.
T. Eagles and En. A. Floss: In. Erg. Chum., vol. 36, 604, 1944; R. K. Cannon, J. Blot. Chum., vol. 152, 401, 1944), and the separation of amino acids using H-type strongly acidic cation exchange resins or type strongly basic anion exchange resins (S. M.
Partridge and R. C. Blimley, Become. J., vol. Sly 628, 19~2).
For industrial isolation of reaction products by ion exchange resins, Japanese Patent Publication No. 5050/1964 discloses a method which comprises adding a polyamide-type polymeric flocculent to a solution containing a saccharine and an amino acid to flocculate and precipitate impurities, and purifying it on an ion exchange resin; and Japanese Patent Publication No. 21105/1973 discloses a method of Jo .
~2~5~69 recovering a mixed amino acid solution including L-tryptophan by a sulfonic acid-type ion exchange resin having a degree of cross linkage represented by a DUB content of not more than 6%.
These prior methods, however, have not proved to be satisfactory for industrial production of amino acids because prior to purification by ion exchange resins, these methods require removal of the used microorganisms from the reaction mixtures by, for example, centrifugal separation, flocculation and precipitation by addition of polymeric flocculants, filtration on an ultrafiltration membrane, etc., and a great deal of labor must go into the removal of the microorganisms.
It is an object of this invention is to provide a method of recovering L-tryptophan effuse-entry from an L-tryptophan-containing reaction mixture produced by using a microorganism.
The object of this invention is achieved by treating the L-tryptophan containing reaction mixture directly with an H type strongly acidic cation exchange resin. The method of this invention enables both the removal of the microorganism used in the reaction and the isolation of the resulting L-tryptophan to be effected simultaneously.
The basic principle of the present invention is that when an L-tryptophan-containing reaction slug mixture containing a microorganism is passed through a layer of an H-type strongly acidic cation exchange resin to adsorb the L-tryptophan on the resin layer and isolate and purify it, the microorganism dissolved or suspended in the reaction mixture is flocculated upon contact with the ion exchange resin and can be easily removed by washing the resin with water. Flocculation of the microorganism occurs presumably because when the microorganism dissolved or suspended in water contacts the H-type strongly acidic cation exchange resin, it is modified by the sulfonic acid group on the ion exchange group The method of this invention is applicable to a reaction mixture containing a microorganism and an L-tryptophan (to be sometimes referred to simply as "amino acid reaction mixture"
hereinafter) obtained during the production of the L-tryptophan utilizing the microorganism.
Examples of such amino acid reaction mixture include an L-tryptophan-containing reaction mixture produced from DL-serine and insole in the presence of Escherichia golf and Pseudomonas putted; an L-tryptophan-containing reaction mixture produced from L-serine and insole in the presence of Escherichia golf; an L-tryptophan-containing reaction mixture produced in the presence of Bacillus subtilis using anthranilic acid as a precursor; an L-tryptophan-containing reaction mixture produced from insole, pyruvic acid and ammonia in the presence of ~erobacter arenas;
and an L-tryptophan containing reaction mixture produced from insole, shrine and glucose using a bacterium of the genus Aerobacterium. These are only illustrative, and the method of I .- -Lowe ._ 4 this invention can be broadly applied to the purification of allL-tryptophan reaction mixtures obtained by utilizing micro-organisms.
Illustrative of the H-type strongly acidic cation exchange resin used in the method of this invention are Lightweight spy (a trade name for a product of Bayer A), Lightweight so 102 (a trade name for a product of Bayer A), Dunn pk-208 (a trade name for a product of Mitsubishi Chemical Co., Ltd.), Dunn Skye (a trade name for a product of Mitsubishi Chemical Co., Ltd.), and Amberlite ZOO (a trade name for a product of Room & Hays Co.).
The amino acid reaction mixture, either as such or if the amino acid precipitates as crystals in water, after diluting it with water until the crystals ~LS~i9 dissolve at room temperature, is passed through a layer of a sulfonic acid type cation exchange resin regenerated to an H-form from its one end to adsorb the amino acid on the ion exchange resin. The micro-S organism in the reaction mixture is modified by contact with the ion exchange resin layer and adheres to the ion exchange resin in the flocculated state.
Then, water is passed at a fixed flow rate through the ion exchange resin layer from the other end of the resin layer (this procedure is called back washing) to remove the flocculated microorganism away from the ion exchange resin layer.
The adsorption of the amino acid, the phlox-lotion of the microorganism and the removal of the flocculated microorganism in the ion exchange resin layer are usually carried out in a column filled with the ion exchange resin. No problem arises, however, even if a method is employed in which after adsorption of the amino acid on the ion exchange resin, the ion exchange resin is discharged into a reaction vessel and subjected to sludge washing with water.
When a column filled with an ion exchange resin is used, the amino acid reaction mixture is caused to flow at a fixed flow rate into the resin column from its top to adsorb the amino acid on the ion exchange resin. At this time, almost all the microorganism contained in the amino acid reaction mixture is modified and flocculated in the resin column and physically held onto the resin. A part of the microorganism flows away from the bottom of the resin column together with the discharged liquor.
When after the above operation, water is passed at a fixed flow rate through the ion exchange resin column from its bottom to perform back washing, the flocculated microorganism adhering to the resin comes afloat and flows away from the upper portion of the column, and can thus be removed efficiently.
The microorganism in the L-tryp-tophan reaction mixture fed into the ion exchange resin layer can be removed virtually completely because at the time of adsorption of the L-tryptophan on the ion exchange resin, a part of the microorganism is removed together with the discharged liquor and at the time of back washing, most of the microorganism in the form of a flocculated mass is removed from the resin layer.
The ion exchange resin having the L-trypto-plan adsorbed thereon is then treated usually with aqueous ammonia to elude the L-tryptophan. By con-cent rating and crystallizing the elude, the desired L-tryptophan can be easily isolated.
The resulting L- tryptophan has a high purity owing to the effect of purification by the ion exchange resin.
By treating an L- tryptophan reaction mixture ~LS~i9 containing a microorganism with an H-type strongly acidic cation exchange resin, the method of this invention makes it possible to simultaneously isolate the L-tryptophan and remove the microorganism which is very difficult to remove industrially by usual methods.
Hence, the method of the invention is of great industrial significance as a method of purifying products obtained by reactions which involve the use of microorganisms.
The following Examples illustrate the method of this invention in greater detail.
Exhume 1 Cells containing Escherichia golf were cult-voted at a pi of 7 and a temperature of 30C with agitation and aeration in the presence of monopotassium phosphate, dipotassium phosphate, ammonium sulfate, calcium chloride, iron sulfate, yeast extract, polyp petunia and other required materials while adding glucose and insole. The final concentration of the cells was 30 to 35 g/liter.
In the same way as above, cells containing Pseudomonas putted were cultivated in the same culture medium as above except that it did not contain insole.
In both cases, the grown cells were collected from the culture broth by using an ordinary supercentri-frugal separator, and obtained as a cream cake having a water content of 75 to 85%.
An aqueous solution composed of 77.3 g of ,. ... ...
I
DL-serine, 10.5 g ox ammonium sulfate and 486 g of water was fed into a reactor, and adjusted to pi 8.5 with 23% aqueous ammonia. Then, 51.2 9 of the cream cake of Escherichia golf cell obtained above and 23.2 g __ _ of the cream cake of Pseudomonas putted cells obtained above were added, and the mixture was well stirred.
Furthermore 392 9 of a Tulane solution containing 78.4 g of insole was added and reacted at 35C for 40 hours.
I The amount of L-tryptophan formed in the reaction mass was analyzed by liquid chromatography, and found to be 129.8 g (yield 95.0~ based on insole).
The reaction mixture was distilled to remove Tulane. Then, the reaction mixture was diluted with water so that the L-tryptophan crystals completely dissolved to a concentration of 1.0% by weight.
On the other hand, 4.86 liters of Lightweight spy (a strongly acidic cation exchange resin) regenerated to an H-form by hydrochloric acid was filled into a column. The above L-tryptophan solution (125 9) was passed through the column from its upper end at a fixed flow rate to adsorb L-tryptophan on the ion exchange resin.
Then, the column was back-washed with 24.9 g of water to wash away the floating flocculated mass of the microbial cells. Thereafter, L-tryptophan was eluded from the resin column by using aqueous ammonia in an amount corresponding to twice the exchange capacity of the ion exchange resin. The equate was heated to 100C to remove and recover ammonia. The residue was cooled to room temperature. The precipi-toted L-tryptophan crystals were separated by filter-lion and dried. L-tryptophan having a purity of 99.8 was obtained in an amount of 1.0 g.
The cell balance by the ion exchange resin treatment was such that I of the cells existed in the waste liquor which passed through the column at the time of adsorption of L-tryptophan and 97% of the cells existed in the effluent at the time of back washing (the cell balance was determined from the weight of the cells which were concentrated to dryness lo and the carbon balance obtained by elemental analysis).
Example 2 L-tryptophan was produced from L-serine and insole in water using a cream cake of Escherichia golf cells cultivated in the same way as in Example 1. To avoid a reduction in the activity of the enzyme by insole, the faction was carried out by adding insole gradually while continuously analyzing its concern-traction so that the insole concentration in water was maintained at 200 Pam or lower. The yield of L-trypto-plan produced was 100~ based on insole and 85% based on L-serine. The final concentration of L-trypto-plan accumulated was 12~ g/liter. The L-tryptophan I
reaction mixture was dehydrated centrifugally to obtain a reaction cream cake containing L-tryptophan and the microbial cells.
The reaction cream cake was treated with Lightweight Skye (H-form) as an ion exchange resin by the same procedure as in Example 1 to remove the cells and isolate L-tryptophan~
The amount of L-tryptophan isolated was 1.1 9 and its purity was 99.9~. The cells in the L-trypto-plan reaction mixture were removed in an amount of
This invention relates to a method of effuse-entry isolating L-tryptophan from a reaction mixture obtained during the production of L-tryptophan my utilizing a microorganism.
Many methods have been reported previously for isolating an L-amino acid by an ion exchange resin from an L-amino acid-containing reaction mixture produced by utilizing a microorganism. For example, there have been known the separation of basic amino acids by carboxylic acid-type cation exchange resins (U. So Patent No. 2,549,378), the separation of acidic amino acids by weakly basic anion exchange resins (D.
T. Eagles and En. A. Floss: In. Erg. Chum., vol. 36, 604, 1944; R. K. Cannon, J. Blot. Chum., vol. 152, 401, 1944), and the separation of amino acids using H-type strongly acidic cation exchange resins or type strongly basic anion exchange resins (S. M.
Partridge and R. C. Blimley, Become. J., vol. Sly 628, 19~2).
For industrial isolation of reaction products by ion exchange resins, Japanese Patent Publication No. 5050/1964 discloses a method which comprises adding a polyamide-type polymeric flocculent to a solution containing a saccharine and an amino acid to flocculate and precipitate impurities, and purifying it on an ion exchange resin; and Japanese Patent Publication No. 21105/1973 discloses a method of Jo .
~2~5~69 recovering a mixed amino acid solution including L-tryptophan by a sulfonic acid-type ion exchange resin having a degree of cross linkage represented by a DUB content of not more than 6%.
These prior methods, however, have not proved to be satisfactory for industrial production of amino acids because prior to purification by ion exchange resins, these methods require removal of the used microorganisms from the reaction mixtures by, for example, centrifugal separation, flocculation and precipitation by addition of polymeric flocculants, filtration on an ultrafiltration membrane, etc., and a great deal of labor must go into the removal of the microorganisms.
It is an object of this invention is to provide a method of recovering L-tryptophan effuse-entry from an L-tryptophan-containing reaction mixture produced by using a microorganism.
The object of this invention is achieved by treating the L-tryptophan containing reaction mixture directly with an H type strongly acidic cation exchange resin. The method of this invention enables both the removal of the microorganism used in the reaction and the isolation of the resulting L-tryptophan to be effected simultaneously.
The basic principle of the present invention is that when an L-tryptophan-containing reaction slug mixture containing a microorganism is passed through a layer of an H-type strongly acidic cation exchange resin to adsorb the L-tryptophan on the resin layer and isolate and purify it, the microorganism dissolved or suspended in the reaction mixture is flocculated upon contact with the ion exchange resin and can be easily removed by washing the resin with water. Flocculation of the microorganism occurs presumably because when the microorganism dissolved or suspended in water contacts the H-type strongly acidic cation exchange resin, it is modified by the sulfonic acid group on the ion exchange group The method of this invention is applicable to a reaction mixture containing a microorganism and an L-tryptophan (to be sometimes referred to simply as "amino acid reaction mixture"
hereinafter) obtained during the production of the L-tryptophan utilizing the microorganism.
Examples of such amino acid reaction mixture include an L-tryptophan-containing reaction mixture produced from DL-serine and insole in the presence of Escherichia golf and Pseudomonas putted; an L-tryptophan-containing reaction mixture produced from L-serine and insole in the presence of Escherichia golf; an L-tryptophan-containing reaction mixture produced in the presence of Bacillus subtilis using anthranilic acid as a precursor; an L-tryptophan-containing reaction mixture produced from insole, pyruvic acid and ammonia in the presence of ~erobacter arenas;
and an L-tryptophan containing reaction mixture produced from insole, shrine and glucose using a bacterium of the genus Aerobacterium. These are only illustrative, and the method of I .- -Lowe ._ 4 this invention can be broadly applied to the purification of allL-tryptophan reaction mixtures obtained by utilizing micro-organisms.
Illustrative of the H-type strongly acidic cation exchange resin used in the method of this invention are Lightweight spy (a trade name for a product of Bayer A), Lightweight so 102 (a trade name for a product of Bayer A), Dunn pk-208 (a trade name for a product of Mitsubishi Chemical Co., Ltd.), Dunn Skye (a trade name for a product of Mitsubishi Chemical Co., Ltd.), and Amberlite ZOO (a trade name for a product of Room & Hays Co.).
The amino acid reaction mixture, either as such or if the amino acid precipitates as crystals in water, after diluting it with water until the crystals ~LS~i9 dissolve at room temperature, is passed through a layer of a sulfonic acid type cation exchange resin regenerated to an H-form from its one end to adsorb the amino acid on the ion exchange resin. The micro-S organism in the reaction mixture is modified by contact with the ion exchange resin layer and adheres to the ion exchange resin in the flocculated state.
Then, water is passed at a fixed flow rate through the ion exchange resin layer from the other end of the resin layer (this procedure is called back washing) to remove the flocculated microorganism away from the ion exchange resin layer.
The adsorption of the amino acid, the phlox-lotion of the microorganism and the removal of the flocculated microorganism in the ion exchange resin layer are usually carried out in a column filled with the ion exchange resin. No problem arises, however, even if a method is employed in which after adsorption of the amino acid on the ion exchange resin, the ion exchange resin is discharged into a reaction vessel and subjected to sludge washing with water.
When a column filled with an ion exchange resin is used, the amino acid reaction mixture is caused to flow at a fixed flow rate into the resin column from its top to adsorb the amino acid on the ion exchange resin. At this time, almost all the microorganism contained in the amino acid reaction mixture is modified and flocculated in the resin column and physically held onto the resin. A part of the microorganism flows away from the bottom of the resin column together with the discharged liquor.
When after the above operation, water is passed at a fixed flow rate through the ion exchange resin column from its bottom to perform back washing, the flocculated microorganism adhering to the resin comes afloat and flows away from the upper portion of the column, and can thus be removed efficiently.
The microorganism in the L-tryp-tophan reaction mixture fed into the ion exchange resin layer can be removed virtually completely because at the time of adsorption of the L-tryptophan on the ion exchange resin, a part of the microorganism is removed together with the discharged liquor and at the time of back washing, most of the microorganism in the form of a flocculated mass is removed from the resin layer.
The ion exchange resin having the L-trypto-plan adsorbed thereon is then treated usually with aqueous ammonia to elude the L-tryptophan. By con-cent rating and crystallizing the elude, the desired L-tryptophan can be easily isolated.
The resulting L- tryptophan has a high purity owing to the effect of purification by the ion exchange resin.
By treating an L- tryptophan reaction mixture ~LS~i9 containing a microorganism with an H-type strongly acidic cation exchange resin, the method of this invention makes it possible to simultaneously isolate the L-tryptophan and remove the microorganism which is very difficult to remove industrially by usual methods.
Hence, the method of the invention is of great industrial significance as a method of purifying products obtained by reactions which involve the use of microorganisms.
The following Examples illustrate the method of this invention in greater detail.
Exhume 1 Cells containing Escherichia golf were cult-voted at a pi of 7 and a temperature of 30C with agitation and aeration in the presence of monopotassium phosphate, dipotassium phosphate, ammonium sulfate, calcium chloride, iron sulfate, yeast extract, polyp petunia and other required materials while adding glucose and insole. The final concentration of the cells was 30 to 35 g/liter.
In the same way as above, cells containing Pseudomonas putted were cultivated in the same culture medium as above except that it did not contain insole.
In both cases, the grown cells were collected from the culture broth by using an ordinary supercentri-frugal separator, and obtained as a cream cake having a water content of 75 to 85%.
An aqueous solution composed of 77.3 g of ,. ... ...
I
DL-serine, 10.5 g ox ammonium sulfate and 486 g of water was fed into a reactor, and adjusted to pi 8.5 with 23% aqueous ammonia. Then, 51.2 9 of the cream cake of Escherichia golf cell obtained above and 23.2 g __ _ of the cream cake of Pseudomonas putted cells obtained above were added, and the mixture was well stirred.
Furthermore 392 9 of a Tulane solution containing 78.4 g of insole was added and reacted at 35C for 40 hours.
I The amount of L-tryptophan formed in the reaction mass was analyzed by liquid chromatography, and found to be 129.8 g (yield 95.0~ based on insole).
The reaction mixture was distilled to remove Tulane. Then, the reaction mixture was diluted with water so that the L-tryptophan crystals completely dissolved to a concentration of 1.0% by weight.
On the other hand, 4.86 liters of Lightweight spy (a strongly acidic cation exchange resin) regenerated to an H-form by hydrochloric acid was filled into a column. The above L-tryptophan solution (125 9) was passed through the column from its upper end at a fixed flow rate to adsorb L-tryptophan on the ion exchange resin.
Then, the column was back-washed with 24.9 g of water to wash away the floating flocculated mass of the microbial cells. Thereafter, L-tryptophan was eluded from the resin column by using aqueous ammonia in an amount corresponding to twice the exchange capacity of the ion exchange resin. The equate was heated to 100C to remove and recover ammonia. The residue was cooled to room temperature. The precipi-toted L-tryptophan crystals were separated by filter-lion and dried. L-tryptophan having a purity of 99.8 was obtained in an amount of 1.0 g.
The cell balance by the ion exchange resin treatment was such that I of the cells existed in the waste liquor which passed through the column at the time of adsorption of L-tryptophan and 97% of the cells existed in the effluent at the time of back washing (the cell balance was determined from the weight of the cells which were concentrated to dryness lo and the carbon balance obtained by elemental analysis).
Example 2 L-tryptophan was produced from L-serine and insole in water using a cream cake of Escherichia golf cells cultivated in the same way as in Example 1. To avoid a reduction in the activity of the enzyme by insole, the faction was carried out by adding insole gradually while continuously analyzing its concern-traction so that the insole concentration in water was maintained at 200 Pam or lower. The yield of L-trypto-plan produced was 100~ based on insole and 85% based on L-serine. The final concentration of L-trypto-plan accumulated was 12~ g/liter. The L-tryptophan I
reaction mixture was dehydrated centrifugally to obtain a reaction cream cake containing L-tryptophan and the microbial cells.
The reaction cream cake was treated with Lightweight Skye (H-form) as an ion exchange resin by the same procedure as in Example 1 to remove the cells and isolate L-tryptophan~
The amount of L-tryptophan isolated was 1.1 9 and its purity was 99.9~. The cells in the L-trypto-plan reaction mixture were removed in an amount of
2.5% at the time of adsorption to the ion exchange resin and 97.5% at the time of back washing of the ion exchange resin column.
Claims (6)
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:
1. A method of isolating L-tryptophan from a reaction mix-ture containing the L-tryptophan and a microorganism which is obtained during the production of the L-tryptophan by utilizing the microorganism; said method comprising treating said reaction mixture with an H-type strongly acidic cation exchange resin while the L-tryptophan is maintained in the dissolved state.
2. A method according to claim 1 wherein the reaction mix-ture is produced from DL-serine and indole in the presence of Escherichia coli and Pseudomonas putida.
3. A method according to claim 1 wherein the reaction mix-ture is produced from L-serine and indole in the presence of Escherichia coli.
4. A method according to claim 1 wherein the reaction mix-ture is produced in the presence of Bacillus subtilis using anthranilic acid as a precursor.
5. A method according to claim 1 wherein the reaction mix-ture is produced from indole, pyruvic acid and ammonia in the presence of Aerobacter aerogenes.
6. A method according to claim 1 wherein the reaction mix-lure is produced from indole, serine and glucose using a bacterium of the genus Aerobacterium.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP13841582A JPS5928484A (en) | 1982-08-11 | 1982-08-11 | Method for isolating l-amino acid |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA1215069A true CA1215069A (en) | 1986-12-09 |
Family
ID=15221422
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA000444165A Expired CA1215069A (en) | 1982-08-11 | 1983-12-22 | Method of isolating l-trytophan |
Country Status (8)
| Country | Link |
|---|---|
| JP (1) | JPS5928484A (en) |
| AU (1) | AU567903B2 (en) |
| CA (1) | CA1215069A (en) |
| CH (1) | CH659827A5 (en) |
| DE (1) | DE3400574A1 (en) |
| FR (1) | FR2557872B1 (en) |
| GB (1) | GB2152030B (en) |
| NL (1) | NL8304496A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8119373B2 (en) | 2006-03-15 | 2012-02-21 | Kyowa Hakko Bio Co., Ltd. | Method for purifying histidine from a cell culture |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4769474A (en) * | 1985-04-10 | 1988-09-06 | Mitsui Toatsu Chemicals, Inc. | Process for purifying tryptophane |
| DE3702689A1 (en) * | 1987-01-30 | 1988-08-11 | Degussa | METHOD FOR ISOLATING L-AMINO ACIDS |
| KR950013858B1 (en) * | 1987-10-12 | 1995-11-17 | 미쓰이 도아쓰 가가쿠 가부시기가이샤 | Process for producing l-tryptophane |
| JPH0325170U (en) * | 1989-07-20 | 1991-03-14 | ||
| EP1995322B1 (en) | 2006-03-15 | 2017-11-22 | Kyowa Hakko Bio Co., Ltd. | Methods for purifying amino acids |
| US20080070282A1 (en) | 2006-08-21 | 2008-03-20 | Samsung Electronics Co., Ltd. | Method and device for obtaining or amplifying nucleic acid from a cell using a nonplanar solid substrate |
| US8158411B2 (en) | 2006-08-21 | 2012-04-17 | Samsung Electronics Co., Ltd. | Method of separating microorganism using nonplanar solid substrate and device for separating microorganism using the same |
| JP6303017B2 (en) * | 2014-01-07 | 2018-03-28 | ノヴァセプ プロセスＮｏｖａｓｅｐ Ｐｒｏｃｅｓｓ | Method for purifying aromatic amino acids |
| CN103772086B (en) * | 2014-01-10 | 2015-04-29 | 国家海洋局第三海洋研究所 | Pretreatment process for preparing fractions of marine microorganism small molecule metabolites |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IT1045451B (en) * | 1966-03-23 | 1980-05-10 | Ajinomoto Kk | METHOD FOR RECOVERING LYSINE FROM FERMENTATION BROTH |
| GB1186952A (en) * | 1967-06-17 | 1970-04-08 | Kyowa Hakko Kogyo Kk | Process for producing L-Tryptophan |
| JPS50127879A (en) * | 1974-03-28 | 1975-10-08 | ||
| JPS5636710A (en) * | 1979-09-04 | 1981-04-10 | Fanuc Ltd | Feed speed command system |
| JPS57174096A (en) * | 1981-04-20 | 1982-10-26 | Ajinomoto Co Inc | Preparation of l-tryptophan by fermentation method |
-
1982
- 1982-08-11 JP JP13841582A patent/JPS5928484A/en active Granted
-
1983
- 1983-12-21 GB GB08334070A patent/GB2152030B/en not_active Expired
- 1983-12-22 CA CA000444165A patent/CA1215069A/en not_active Expired
- 1983-12-23 AU AU22840/83A patent/AU567903B2/en not_active Ceased
- 1983-12-30 NL NL8304496A patent/NL8304496A/en not_active Application Discontinuation
-
1984
- 1984-01-06 CH CH8884A patent/CH659827A5/en not_active IP Right Cessation
- 1984-01-10 FR FR8400302A patent/FR2557872B1/en not_active Expired
- 1984-01-10 DE DE19843400574 patent/DE3400574A1/en active Granted
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8119373B2 (en) | 2006-03-15 | 2012-02-21 | Kyowa Hakko Bio Co., Ltd. | Method for purifying histidine from a cell culture |
Also Published As
| Publication number | Publication date |
|---|---|
| GB2152030A (en) | 1985-07-31 |
| GB8334070D0 (en) | 1984-02-01 |
| AU567903B2 (en) | 1987-12-10 |
| NL8304496A (en) | 1985-07-16 |
| DE3400574A1 (en) | 1985-07-18 |
| JPS5928484A (en) | 1984-02-15 |
| FR2557872B1 (en) | 1987-07-10 |
| CH659827A5 (en) | 1987-02-27 |
| GB2152030B (en) | 1987-08-19 |
| JPH0453509B2 (en) | 1992-08-26 |
| FR2557872A1 (en) | 1985-07-12 |
| DE3400574C2 (en) | 1987-08-06 |
| AU2284083A (en) | 1985-06-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US5017480A (en) | Process for recovering L-amino acid from fermentation liquors | |
| JP2664648B2 (en) | Method for producing L-aspartic acid | |
| CA1215069A (en) | Method of isolating l-trytophan | |
| CA1267652A (en) | Process for purifying tryptophan | |
| US5532148A (en) | Process for producing of citric acid and monovalent citrate salts | |
| HU202282B (en) | Process for separating 2-keto-l-gulonic acid from fermentation juice | |
| US6072083A (en) | Method for purifying branched chain amino acids | |
| JP2668956B2 (en) | Method for purifying L-glutamine | |
| JP4361641B2 (en) | Separation and recovery method of optically active amino acid and optically active amino acid amide | |
| CN1125037C (en) | Process for producing glutamic acid | |
| JP2001258583A (en) | Method for purifying shikimic acid | |
| US4997754A (en) | Process for recovering L-amino acids from fermentation liquors containing them | |
| JPH0739385A (en) | Production of l-3,4-dihydroxyphenylalanine | |
| CA1274250A (en) | Purification and recovery of phenylalanine with calcium salts | |
| JP5092743B2 (en) | Separation and recovery method of optically active amino acid amide | |
| KR860001821B1 (en) | Method of isolating l-amino acids | |
| US3381027A (en) | Preparation and recovery of methyl 2-ketogluconate | |
| SK176398A3 (en) | Process for the production of l-aspartic acid | |
| JPS61234789A (en) | Recovery of indole from triptophane | |
| JP2721536B2 (en) | Method for obtaining D-β-hydroxy amino acid | |
| JP2804005B2 (en) | Method for producing L-aspartic acid | |
| KR960006578B1 (en) | Separating method microorganisms from amino acid and nucleotide fermenting liquid | |
| JPH07308195A (en) | Production of l-aspartic acid | |
| US3336374A (en) | Recovery of l-glutamic acid | |
| JPH0489479A (en) | Recovery of optically active tryptophan |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| MKEX | Expiry |