KR101372811B1 - Tryptophan purification devices and methods - Google Patents

Abstract

A tryptophan purification device is disclosed. The disclosed tryptophan purification device comprises: a solid purification filter for filtering solids included in a tryptophan solution and passing only the solution by passing the tryptophan solution through the filter; a storage tank for storing the tryptophan solution which has passed through the solid purification filter; a heating jacket installed outside the storage tank to control the temperature inside the storage tank; an air compressor for suctioning the tryptophan solution in the storage tank and transferring the solution to the next location; a continuous reactor for inputting the tryptophan solution transferred by the air compressor together with sulfuric acid transferred via a separate route, and continuously generating a reaction product by stirring the input solutions; a dehydrator connected to the discharge port of the continuous reactor to separate filtrate from the reaction product in the form of slurry discharged from the continuous reactor; and a drier for drying the solid components separated by the dehydrator.

Description

Tryptophan purification devices and methods

The present invention relates to a tryptophan purification apparatus and method.

Especially, tryptophan is dissolved in an acid solution or basic solution, and then the opposite solution is added to neutralize the pH and precipitated. In addition, it can be continuously produced using a continuous reactor, and more than three times more than conventional methods. The present invention relates to a tryptophan refining apparatus and method which has high speed, improved recovery and purity, and is not easily broken by improving the density of particles as the polymer material is mixed with an additive.

Tryptophan is one of the essential amino acids and is a useful substance used as feed additives, pharmaceuticals, and health foods.

As a method for producing tryptophan, there is a method (fermentation method) using bacterial cells using glucose or the like as a main starting material. However, crude crystals of tryptophan obtained by this conventional method contain impurities such as proteins and coloring substances. Since these impurities exhibit high hydrophobicity and are therefore incorporated between the crystal surface or the crystals of tryptophan, it is difficult to remove them in any amount, for example by conventional recrystallization.

Therefore, such tryptophan is difficult to pass the standard specification (transmittance (%) = 95 or more, λ = 430 nm) for use as a source of high purity products, such as pharmaceutical supplies.

The prior art includes the method described in Japanese Patent Laid-Open No. 895/1983. This method combines treatment with a non-polar highly porous resin and filtration with an ultrafiltration membrane to remove reaction inhibitors contained and accumulated in the reaction solution of tryptophan, or the recovered solution and the mother liquor for purification.

Another method is characterized by adjusting the pH of the reaction solution of tryptophan to alkalinity and crystallizing it in the presence of a lower alcohol or ketone under a neutralization reaction (see Japanese Patent Laid-Open No. 39857/1984). number).

In a third method, the enzyme reactant of tryptophan is heated to 95-100 ° C. under acidic pH values in the presence of activated carbon, the liquid solution is removed from the solids, and the filtered solution containing tryptophan is a non-polar highly porous resin. And a lower aliphatic alcohol is added to the solution (see Japanese Patent Laid-Open No. 126070/1986).

There is a need to develop a purification method to easily obtain a high purity colorless tryptophan having a transmittance (%) of 95% or more from the tryptophan-prepared reaction solution. As a result of various tests, the molecular weight of the impurity reducing the permeability is 500 to 5,000, and the method using the ultrafiltration membrane according to the method described in JP 895/1983 can only remove impurities at a fixed removal rate. It has been found that excessive loads on the resin may occur, thereby shortening the regeneration cycle of the nonpolar highly porous resin.

In addition, according to only the method described in Japanese Patent Laid-Open No. 39857/1984, it is difficult to obtain crystals having a low degree of removal of impurities and having a transmittance of 95% or more.

Further, in the case of the method described in Japanese Patent Laid-Open No. 126070/1986, tryptophan, especially when the reaction solution prepared by the fermentation method containing a large amount of impurities, is heated to 95 to 100 ° C in the presence of activated carbon, Since it contains an unstable indole ring in its structure, there is a problem of increasing decomposition and coloring materials.

The present invention takes a method of dissolving tryptophan in an acid solution or a basic solution and then adding the opposite solution to neutralize and precipitate the pH, and continuously producing it using a continuous reactor, and more than three times compared to the conventional method. It is to provide a tryptophan refining apparatus having a fast production speed, improved recovery and purity, and improves the density of particles as the polymer material is mixed with an additive, thereby not easily breaking.

In addition, the present invention provides a method using the tryptophan purification apparatus.

According to a first aspect of the present invention for achieving the above object, as a purification apparatus used in the tryptophan purification method described above, the tryptophan purification apparatus,

A solid filtration filter passing through the tryptophan solution to filter the solids contained in the tryptophan solution and passing only the solution; A storage tank in which the tryptophan solution passing through the solid filtration filter is stored; A heating jacket installed outside the storage tank to control a temperature of the storage tank; An air compressor which sucks the tryptophan solution in the storage tank and sends it to the next position; A continuous reactor in which the tryptophan solution sent out by the air compressor and sulfuric acid sent out through a separate path are added together, and the reacted solutions are stirred to continuously generate a reactant; A dehydrator connected to the discharge port of the continuous reactor to separate the filtrate from the slurry in the form of a reactant discharged from the continuous reactor; And a dryer for drying the solid component separated from the dehydrator.

In addition, according to the first aspect of the present invention, a temperature sensor is installed in the storage tank, so that the temperature of the solution stored in the storage tank can be detected in real time.

On the other hand, according to the second aspect of the present invention,

A solids removing step (S10) of removing the solids having a predetermined size or more by passing the filtration filter when the tryptophan solution is stored in the storage tank; A hydrogen ion index adjusting step (S20) of adjusting NaH in a storage tank to adjust a hydrogen ion index (pH) of the tryptophan solution; A temperature control step (S30) of installing a heating jacket on the outside of the storage tank to maintain a constant temperature of the tryptophan solution in the storage tank; Density enhancement step (S40) of adding a polymer material to the tryptophan solution in the storage tank to improve the particle density of the tryptophan solution; Injecting the tryptophan solution in the storage tank into a continuous reactor (S50); Adding an acid solution to the tryptophan solution introduced into the continuous reactor to neutralize the hydrogen ion index to precipitate particulate crystals (S50); Dehydrating the precipitated particulate crystals using a dehydrator to separate solid-liquid (S60); And a tryptophan purification method comprising the step of drying the solid component with hot air (S70).

In addition, according to the second aspect of the present invention, the mesh size of the filtration filter in the solids removal step is characterized in that 0.1 ~ 0.5㎛.

Further, according to the second aspect of the present invention, the hydrogen ion index (pH) in the hydrogen ion index adjustment step is characterized in that 11 or more.

In addition, according to the second aspect of the present invention, the polymer material in the density improving step is characterized in that the polyvinyl alcohol (PVA).

In the present invention, the recovery rate of tryptophan is improved by about 10% by the neutralization crystallization process and the continuous reactor, the production rate is improved by three times or more, and the purity is also improved to 99.9% or more by the continuous production.

In addition, the present invention improves the density of the particles by adding a high molecular material and does not easily break.

1 is a flow chart for a tryptophan purification method according to the invention.
2 to 4 are photographs showing the change of crystals according to the sulfuric acid injection rate in the tryptophan purification method according to the present invention.
5 is a graph of the particle size distribution according to the reaction time in the tryptophan purification method according to the present invention.
6 to 9 is a tryptophan purification method according to the present invention, the photo showing the crystal shape of tryptophan according to the reaction time (Fig. 6 1 hour / Fig. 6 6 hours / Fig. 8 26 hours / Fig. 9 52 hours to be).
10 is a graph showing the change in particle size according to the tryptophan dissolution concentration in the tryptophan purification method according to the present invention.
11 is a graph showing a change in recovery rate according to the tryptophan dissolution concentration in the tryptophan purification method according to the present invention.
12 is a graph showing a recovery rate according to the tryptophan dissolution concentration in the tryptophan purification method according to the present invention.
13 to 14 is a tryptophan purification method according to the invention, the photo showing the change in particle shape according to the stirring speed (Fig. 13 is 100rpm / Fig. 14 is 300rpm).
15 is a graph showing the particle size change according to the stirring speed in the tryptophan purification method according to the present invention.
16 is a block diagram of a tryptophan purification apparatus according to the present invention.

Hereinafter, embodiments according to the present invention are disclosed.

In the disclosed embodiments, the same reference numerals are used to designate the same or similar elements, and additional descriptions that may be used to limit or interpret the meaning of the invention may be omitted from the description of embodiments of the present invention.

Prior to the description of the present invention, in the context of the environment in which the singular / plural is not clearly distinguished in the use of the Korean language and the conventional usage environment of the term in the field, although the singular expression is described in the present specification, And is used in a sense to include plural expressions unless it is contrary to the interpretation or expressly contradicts otherwise. It is also to be understood that the word "comprise," "comprising," "comprising," "comprising," and the like, which may be described orended herein, Should not be excluded.

As used herein, the term " composition " of the " binder composition " in the present invention refers to the term " composition " as used in the present invention unless otherwise specified or contrary to the entire contents of the invention. It is used in the meaning of.

Tryptophan purification method to be disclosed in the present invention follows the following procedure (see Figure 1).

Step 1, solids removal step (S10).

The solids removing step is a process of filtering solids having a predetermined size or more by passing through a filtration filter before storing the tryptophan solution in a storage tank.

Here, the mesh size (or pore size) of the filtration filter in the solids removing step is preferably having a size of 0.1 ~ 0.5㎛. Therefore, dissolved solids of 0.5 μm or more contained in the tryptophan solution are filtered so that the tryptophan solution includes only fine solids of 0.5 μm or less.

Step 2, the hydrogen ion index adjustment step (S20).

Hydrogen ion index adjustment step is to adjust the hydrogen ion index (pH) of tryptophan solution by introducing NaOH (caustic soda) from the NaOH (caustic soda) supply tank connected to the storage tank in the storage tank, where hydrogen The ion index (pH) is preferably 11 or more or 3 or less.

Step 3, the temperature control step (S30).

In the temperature control step, a heating jacket is installed outside the storage tank to maintain the tryptophan solution temperature in the storage tank at a constant temperature.

Four steps, the density improvement step (S40).

The density enhancing step is a step of improving the particle density of the tryptophan solution by adding a polymer material to the tryptophan solution in the storage tank. This step is accomplished using a continuous reactor.

Here, the polymer material may be any one selected from PVA (polyvinyl alcohol), Carrageenan, alginic acid, HPC, gellatin. As can be seen in Figure 4, for example, in both the case of applying PVA and gellatin, it can be seen that the growth as a large crystal over time, a high growth rate is confirmed when compared to the crystal of tryptophan without the polymer added As the content of acid solution (especially sulfuric acid), which will be explained in the next six steps, increases, it grows into relatively large crystals.

In addition, the addition of the polymer material causes the particles of the tryptophan solution to grow to a size of about 300 μm or more. In this case, as a result of the sonication, the strength of the particles is weak and easily broken. However, when the molecular weight of the polymer material is adjusted, the strength of the particles is improved when the molecular weight is small.

Step 5 is the input step (S50).

The injecting step is a process of injecting the tryptophan solution in the storage tank into the continuous reactor, and the injecting process may be automatically performed by an air compressor or the like. The configuration of the continuous reactor will be described later.

Step 6 is a crystal precipitation step (S60).

In the crystal precipitation step, an acid solution (particularly sulfuric acid) is added to the tryptophan solution introduced into the continuous reactor to neutralize the hydrogen ion index to precipitate particulate crystals.

That is, the tryptophan solution is well dissolved in an acid and a base solution as an amphoteric substance.The tryptophan solution is dissolved in an acid solution or a basic solution, and then the opposite solution is added (for example, an acid solution is injected after dissolving in a sodium hydroxide solution). The pH is neutralized to precipitate crystals.

Step 7 is a solid-liquid separation step (S70).

The solid-liquid separation step is a process of separating solid-liquid by dehydrating particulate crystals precipitated in the continuous reactor by a dehydrator.

That is, the obtained tryptophan in a high moisture content state is put into a centrifugal carburettor, and the dehydrator is operated to lower the water content to 60% or less. In order to calculate an appropriate dehydration time, dehydration is performed for an arbitrary time. It is necessary to collect the dehydrated sludge tryptophan, and measure the moisture content using a moisture content meter, and adjust the dehydration time according to the measurement result.

The preferred embodiment obtained in the above process is the rotational speed of the dehydrator is 5,000 ~ 12,000rpm, the dehydration time is 50 ~ 60 minutes. When rotating at a lower speed than 500rpm, the dehydration time is too long, and when rotating at a higher speed than 12,000rpm, there is no significant difference in the dehydration rate as compared to when rotating at a smaller rpm.

Eighth step is a drying step (S80).

In the drying step, since the dehydrated tryptophan sludge cake is still very high in water content, it should be powdered by lowering the water content to 10% or less by hot air drying. Here, the process of washing the sludge cake before hot air drying the sludge cake, and separating the wash water by a centrifuge may be added. Through this powdering process, tryptophan of high purity is obtained.

(Experiment 1)

Sulfuric acid injection rate change experiment

(Experimental conditions)

Tryptophan concentration: 200 g / L

NaOH concentration: 5 mol / L

Initial pH: 14

Sulfuric acid concentration: 30%

Stirring Speed: 300 rpm

-Reaction temperature: 25

Sulfuric acid injection rate: 0.5 mL / min, 2 mL / min

(Experiment result)

The slower the rate of sulfuric acid injection, the smaller the crystal tends to be.The longer the reaction time, the smaller the particle size, so the reaction time should be shorter.

Particle size Cloud point Final pH
PSA (μm) Microscope time pH One 48.64 40 to 45 Immediately - 7.6 2 31 20 ~ 30 37min 9.26 7.7 3 12.49 10-20 110min 10.3 7.6

For reference, FIGS. 2 to 4 are micrographs for cases 1, 2, and 3 of Table 1 above.

(Experiment 2)

Response time confirmation experiment

(Experimental conditions)

Tryptophan concentration: 100 g / L

NaOH concentration: 5 mol / L

Initial pH: 14

Sulfuric acid concentration: 30%

Stirring Speed: 300 rpm

-Reaction temperature: 25

Sulfuric acid injection rate and rate: 100 mL & 16.7 mL / min

-Other conditions: Long time stirring after neutralization to pH 7

(Experiment result)

In the above experimental results, it was possible to produce tryptophan with large particles even though the reaction was performed for a short time, but the results showed that the small crystals were weakly bound and were easily broken during the short time stirring. As shown in FIG. 5 and FIGS. 6 to 9, as the reaction time increases, large crystals tend to increase while the medium crystals decrease. However, it shows uniform particle distribution at reaction time of about 18 hours.

The recovery rate was about 95% or more without significant change depending on the reaction time, and it was confirmed that tryptophan precipitated in a short time.

(Experiment 3)

Tryptophan Concentration Change Experiment

(Experimental conditions)

 NaOH concentration: 5 mol / L

 Tryptophan concentration: 10, 100, 200 g / L

 Sulfuric acid concentration: 30%

 -Stirring speed: 100, 300 rpm

 -Reaction temperature: 25

 pH: 7

Reaction time: 6 h

(Experiment result)

As shown in FIG. 10, the lower the concentration of tryptophan, the larger the particle size. As shown in FIG. 11, the purity was about 99% or more under all conditions, and as shown in FIG. 12, the recovery was about 75% under all conditions. Reducing the concentration of tryptophan is one of the methods of suppressing nucleation, and it was found that this method is suitable. As the nucleation occurs less, the dissolved tryptophan is attached to the nucleus, and crystal growth is believed to have occurred.

(Experiment 4)

Stirring  Speed change experiment

(Experimental conditions)

 NaOH concentration: 5 mol / L

 Tryptophan concentration: 10 g / L

 Sulfuric acid concentration: 30%

 -Stirring speed: 100, 300 rpm

 -Reaction temperature: 25

 pH: 7

 Reaction time: 6 h

(Experiment result)

13 to 15, it was found that the particle size increases as the stirring speed decreases. This material is a plate-like material with a weak bond strength of particles. As a result of analysis, the height of the plate was 1 μm or less, so that the crystals were easily formed.

Hereinafter, a tryptophan purification apparatus for performing the above tryptophan purification method will be described.

Tryptophan purification apparatus according to the present invention, as shown in Figure 16, the solid filtration filter 10, storage tank 20, heating jacket 30, air compressor 40, continuous reactor 50, dehydrator 60 ) And a dryer 70, and a second storage tank 80 for supplying an acid solution.

The solid filtration filter 10 passes through the tryptophan solution so that the solids contained in the tryptophan solution are filtered and only pass through the solution. Here, the solid filtration filter 10 has a mesh size (or pore) of 0.1 μm or more, preferably 0.1 to 0.5 μm, so as to pass through the tryptophan solution, all solids of 0.5 μm or more contained in the tryptophan solution are filtered out. It is removed and only the fine solids below it are passed through, resulting in a first purification step.

The storage tank 20 functions to store the tryptophan solution that has passed through the solid filtration filter. At this time, the storage tank 20 has a temperature sensor 21 is installed to be able to detect the temperature of the tryptophan solution stored in the storage tank in real time.

The heating jacket 30 is installed outside the storage tank 20 to control the temperature inside the storage tank. The heating jacket 30 is installed so that a portion of the storage tank 20 is submerged in a state in which the heat medium is filled, and controls the tryptophan solution in the storage tank 20 to an appropriate temperature as the heat medium is heated by a heat source. You can do it.
On the other hand, the present invention is provided with a NaOH (caustic soda) supply tank (not shown) for supplying NaOH (caustic soda) to the storage tank 20. The NaOH (caustic soda) supply tank is connected to the storage tank and supplies NaOH (caustic soda) stored therein so that the hydrogen ion index (pH) of the tryptophan solution becomes 11 or more or 3 or less.

The air compressor 40 functions to suck the tryptophan solution in the storage tank 20 and to deliver it to the next position, and a pump may be installed in place of the air compressor.

The continuous reactor 50 is supplied with the tryptophan solution sent out by the air compressor 40 and the sulfuric acid sent from the second storage tank 80 together with a separate path, that is, as shown in the drawing, and the reactants are stirred by stirring the injected solutions. It serves to continuously produce

The continuous reactor is based on Patent Registration No. 10-1092337 registered by the person, so a separate description of the configuration is omitted.

The dehydrator 60 is connected to the discharge port of the continuous reactor to serve to separate the filtrate from the slurry in the form of a slurry discharged from the continuous reactor, centrifugal separation can be applied.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is to be understood that the invention is not limited to the disclosed exemplary embodiments, but, on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the invention.

10: solid filtration filter 20: storage tank
30: heating jacket 40: air compressor
50: continuous reactor 60: dehydrator
70: dryer

Claims (6)

A solid filtration filter having a mesh size of 0.1 to 0.5 μm and passing through the tryptophan solution to filter dissolved solids of 0.5 μm or more contained in the tryptophan solution, and passing only the solution;
A storage tank for supplying and storing the tryptophan solution that has passed through the solid filtration filter;
A NaOH (caustic soda) supply tank connected to the storage tank and supplying NaOH (caustic soda) stored therein to the storage tank to adjust the hydrogen ion index (pH) of the tryptophan solution;
A heating jacket installed outside the storage tank to allow a portion of the storage tank to be submerged and to control a temperature inside the storage tank;
An air compressor which sucks the tryptophan solution in the storage tank and sends it to the next position;
Tryptophan solution sent out by the air compressor, PVA, carrageenan (Carrageenan), alginic acid (alginic acid), a continuous polymer to improve the density of the tryptophan solution by agitation any one of the polymer material selected from (HPC, gelatin) Reactor;
A second storage tank connected to the continuous reactor to supply an acid solution, and neutralize the hydrogen ion index of the tryptophan solution through the acid solution to precipitate particulate crystals;
A dehydrator connected to the discharge port of the continuous reactor and separating the filtrate from the slurry in the form of a reactant supplied from the continuous reactor; And
A dryer for drying the solid component separated from the dehydrator;
Tryptophan purification device comprising a.
The method of claim 1,
The storage tank is equipped with a temperature sensor, tryptophan refining device, characterized in that to detect the temperature of the solution stored in the storage tank in real time.
As a tryptophan purification method using the tryptophan purification apparatus according to claim 1 or 2,
A solids removing step (S10) of removing the solids having a predetermined size or more by passing a filtration filter when storing the tryptophan solution in a storage tank;
A hydrogen ion index adjusting step (S20) of adjusting NaH in a storage tank to adjust a hydrogen ion index (pH) of the tryptophan solution;
A temperature control step (S30) of installing a heating jacket on the outside of the storage tank to maintain a constant temperature of the tryptophan solution in the storage tank;
Density enhancement step (S40) of adding a polymer material to the tryptophan solution in the storage tank to improve the particle density of the tryptophan solution;
Injecting step (S50) for injecting the tryptophan solution in the storage tank into a continuous reactor;
A crystal precipitation step (S60) in which an acid solution is added to the tryptophan solution introduced into the continuous reactor to neutralize the hydrogen ion index to precipitate particulate crystals (S60);
A solid-liquid separation step (S70) of separating the solid-liquid by dehydrating the precipitated particulate crystals by a dehydrator; And
A drying step of drying the solid component with hot air (S80);
Tryptophan purification method comprising a.
The method of claim 3,
Tryptophan purification method characterized in that the mesh size of the filtration filter in the solids removal step is 0.1㎛ or more.
The method of claim 3,
Tryptophan purification method characterized in that the hydrogen ion index (pH) in the hydrogen ion index adjustment step is 11 or more or less than 3.
The method of claim 3,
Tryptophan purification method characterized in that the polymer material in the step of increasing the density is polyvinyl alcohol (PVA).

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