WO2021107451A1 - 암모니아의 지속 가능한 순환이 가능한 분지쇄 아미노산의 결정화 방법 - Google Patents
암모니아의 지속 가능한 순환이 가능한 분지쇄 아미노산의 결정화 방법 Download PDFInfo
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- WO2021107451A1 WO2021107451A1 PCT/KR2020/015633 KR2020015633W WO2021107451A1 WO 2021107451 A1 WO2021107451 A1 WO 2021107451A1 KR 2020015633 W KR2020015633 W KR 2020015633W WO 2021107451 A1 WO2021107451 A1 WO 2021107451A1
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- Prior art keywords
- chain amino
- amino acid
- branched chain
- ammonia
- branched
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Images
Classifications
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C229/00—Compounds containing amino and carboxyl groups bound to the same carbon skeleton
- C07C229/02—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton
- C07C229/04—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated
- C07C229/06—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton
- C07C229/08—Compounds containing amino and carboxyl groups bound to the same carbon skeleton having amino and carboxyl groups bound to acyclic carbon atoms of the same carbon skeleton the carbon skeleton being acyclic and saturated having only one amino and one carboxyl group bound to the carbon skeleton the nitrogen atom of the amino group being further bound to hydrogen atoms
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
- C07B2200/00—Indexing scheme relating to specific properties of organic compounds
- C07B2200/13—Crystalline forms, e.g. polymorphs
Definitions
- the present application relates to a method for crystallizing branched chain amino acids capable of sustainable circulation of ammonia.
- Branched chain amino acid is an amino acid having a branched aliphatic side-chain.
- 20 proteinogenic amino acids leucine, isoleucine ( isoleucine), and valine.
- branched-chain amino acids play a very important role in building muscle, so they are treated as essential substances for people who train mainly for anaerobic exercise.
- branched-chain amino acids are one of the most used health supplements after protein supplements, along with glutamine and creatine.
- the branched-chain amino acid has a lower solubility in pure water than other hydrophilic amino acids due to the hydrophobic aliphatic side chain.
- an increase in the concentration of the target material in the fermentation broth due to the development of a new fermentation technology is considered inevitable. That is, in the industrial fermentation process, when branched-chain amino acids having a concentration higher than the solubility of branched-chain amino acids are included in the fermentation broth, amino acid crystals will be formed in the prepared fermentation broth, which inhibits the pretreatment of the fermentation broth, or the recovery rate of branched-chain amino acid crystals may act as a reducing factor. Therefore, in order to solve this problem, there is a need for a new technology capable of enhancing the solubility of branched-chain amino acids in fermentation broth or suspension.
- Patent Document 1 Republic of Korea Patent No. 10-1736654
- Another object of the present application is to provide a branched chain amino acid crystal produced by the above method.
- the present application in one aspect, (a) mixing the reaction solution containing the branched chain amino acid crystals with ammonia to obtain a solution in which the branched chain amino acid crystals are dissolved; (b) crystallizing the obtained solution to obtain a concentrated solution containing branched chain amino acid crystals; (c) obtaining a mixed gas including water vapor and ammonia, generated in the crystallization process; And (d) reusing the ammonia derived from the obtained mixed gas as the ammonia of the step (a); including, wherein the steps (b) and (c) are carried out simultaneously or sequentially, branched chain A method for crystallizing amino acids is provided.
- the method of the present application may include a step of mixing a reaction solution containing branched chain amino acid crystals with ammonia to obtain a solution in which branched chain amino acid crystals are dissolved.
- branched-chain amino acid refers to an amino acid having a branched aliphatic side chain, and may be at least one amino acid selected from the group consisting of leucine, isoleucine, and valine, for example, For example, it may be at least one amino acid selected from the group consisting of L-leucine, L-isoleucine, and L-valine.
- mixing with ammonia may increase the pH of the reaction solution containing the branched chain amino acid crystals and increase the solubility of the branched chain amino acid in the reaction solution.
- separate ammonia for example, ammonia water
- step (c) It may be mixing ammonia derived from the mixed gas.
- the reaction solution containing the branched-chain amino acid crystals may be in the form of a suspension in a solution in which the branched-chain amino acids are supersaturated, for example, it may include a solution or a fermentation broth containing the branched-chain amino acid crystals.
- the solution containing the branched-chain amino acid crystals may be a mixture of branched-chain amino acid crystals and distilled water, and the fermentation broth containing the branched-chain amino acid crystals may be obtained by culturing a microorganism producing branched-chain amino acids in a medium. have.
- the microorganism producing the branched chain amino acids is not particularly limited as long as it is a microorganism having branched chain amino acid production ability, for example, Corynebacterium , or Escherichia . It may be of the genus, specifically, Coryne Bacterium glutamicum ( Corynebacterium glutamicum ) It may be a strain or a mutant thereof, and in one embodiment, it may be a Corynebacterium glutamicum mutant having an accession number KCCM11662P, KCCM11248P or KCCM11336P.
- the term "cultivation” means growing microorganisms in an artificially controlled environment.
- the method for producing branched-chain amino acids using a microorganism having branched-chain amino acid-producing ability may be performed using a method widely known in the art. Specifically, the culture may be continuously cultured in a batch process, an injection batch or a repeated fed batch process, but is not limited thereto.
- culture media for Corynebacterium sp. strains are known (eg, Manual of Methods for General Bacteriology. American Society for Bacteriology. Washington D.C., USA, 1981).
- Sugar sources that can be used include sugars and carbohydrates such as glucose, saccharose, lactose, fructose, maltose, starch, cellulose, oils and fats such as soybean oil, sunflower oil, castor oil, coconut oil, palmitic acid, Fatty acids such as stearic acid and linoleic acid, alcohols such as glycerol and ethanol, and organic acids such as acetic acid may be included.
- Nitrogen sources that may be used include peptone, yeast extract, broth, malt extract, corn steep liquor, soybean wheat and urea or inorganic compounds such as ammonium sulfate, ammonium chloride, ammonium phosphate, ammonium carbonate and ammonium nitrate.
- the nitrogen source may also be used individually or as a mixture, but is not limited thereto.
- the phosphorus that may be used may include potassium dihydrogen phosphate or dipotassium hydrogen phosphate or the corresponding sodium-containing salt.
- the culture medium may contain a metal salt such as magnesium sulfate or iron sulfate necessary for growth. Additionally, essential growth substances such as amino acids and vitamins may be used. In addition, precursors suitable for the culture medium may be used. The above-mentioned raw materials may be added batchwise or continuously by a method suitable for the culture during the culture process. However, it is not limited thereto.
- the pH of the solution in which the branched chain amino acid crystal obtained in step (a) is dissolved may be 9 to 12.
- the pH of the branched chain amino acid solution is, for example, 9 to 11.5, 9 to 11, 9 to 10.5, 9 to 10.0, 9, to 9.5, 9.5 to 12.0, 9.5 to 11.5, 9.5 to 11.0, or 9.5 to 10.5 , 9.5 to 10, 10 to 12.0, 10 to 11.5, 10 to 11.0, or may be in the range of 10 to 10.5, it can be appropriately adjusted according to the type of amino acid, other reaction conditions, etc.
- the solution in which the branched chain amino acid crystals are dissolved may be changed to a transparent solution in suspension as the branched chain amino acid crystals are dissolved.
- the method of the present application may include crystallizing a solution in which the obtained branched-chain amino acid crystals are dissolved to obtain a concentrate containing the branched-chain amino acid crystals.
- crystallization of the branched chain amino acid solution may be applied to a conventional technique known in the art.
- the crystallization may be accompanied by evaporation of the solvent, and the temperature during the crystallization process is 60 to 90 °C, 60 to 85 °C, 60 to 80 °C, 60 to 75 °C, 60 to 70 °C, 60 to 65 °C, 70 to 90 °C, 70 to 85 °C, 70 to 80 °C, 70 to 75 °C, 75 to 90 °C, 75 to 85 °C, or 75 to 80 °C.
- the crystallization may be performed by a crystallization apparatus without limitation in its configuration and structure, as long as the object of the present application can be achieved.
- the concentrate contains a high concentration of branched chain amino acids, and the concentration of the branched chain amino acids is, for example, 40 to 800 g/L, 55 to 750 g/L, 70 to 700 g/L, 85 to 650 g/L, 100 to It may be 600 g/L, 115 to 550 g/L, 130 to 500 g/L, 145 to 450 g/L, 160 to 400 g/L, 175 to 350 g/L, or 190 to 300 g/L, but this is on the scale of the industrialization process. The enemy can be changed accordingly.
- the branched chain amino acid concentration of the concentrate is 1.0 to 2.0 times, 1.0 to 1.8 times, 1.0 to 1.7 times, 1.0 compared to the branched chain amino acid concentration of the reaction solution comprising the branched chain amino acid crystal of step (a) to 1.6 times, 1.2 to 2.0 times, 1.2 to 1.8 times, 1.2 to 1.7 times, 1.2 to 1.6 times, 1.3 to 2.0 times, 1.3 to 1.8 times, 1.3 to 1.7 times, 1.3 to 1.6 times, 1.4 to 2.0 times, 1.4 to 1.8 times, 1.4 to 1.7 times, or 1.4 to 1.6 times, but is not limited thereto.
- the concentrate may be changed to a suspension form due to increased turbidity of the solution as branched chain amino acid crystals are formed again by crystallization of a transparent branched chain amino acid solution.
- the method of the present application may include obtaining a mixed gas including water vapor and ammonia generated in the crystallization process.
- the step may be carried out simultaneously or sequentially with the step of obtaining a concentrate comprising the aforementioned branched chain amino acid crystals, for example, if the above step can be performed, the composition and structure It can be carried out by any ammonia recovery device without limitation.
- the step may be carried out until the pH of the solution in which the branched chain amino acid crystal of step (b) is dissolved becomes 5.5 to 8.0, and specifically, the pH of the solution 5.5 to 7.5, 5.5 to 7.0, 5.5 to 6.5, 6.0 to 8.0, 6.0 to 7.5, 6.0 to 7.0, 6.0 to 6.5, 6.5 to 8.0, 6.5 to 7.5, 6.5 to 7.0, or 7.0.
- the pH of the solution 5.5 to 7.5, 5.5 to 7.0, 5.5 to 6.5, 6.0 to 8.0, 6.0 to 7.5, 6.0 to 7.0, 6.0 to 6.5, 6.5 to 8.0, 6.5 to 7.5, 6.5 to 7.0, or 7.0.
- the mixed gas may be a gas in a vapor state in which water vapor and ammonia are mixed, and the composition ratio of the mixed gas is dominantly influenced by the gas-liquid equilibrium system of water-ammonia-branched chain ammonia. It may have various distributions depending on the internal temperature and pressure conditions.
- the method of the present application may include reusing ammonia derived from the obtained mixed gas as ammonia in step (a).
- the step may be to reuse the ammonia in a vapor state, or to reuse the vapor in a compressed or condensed liquid state.
- it may further include the step of condensing or compressing the obtained mixed gas to convert ammonia in the mixed gas to a liquid state, for example, condensing the mixed gas using a cooler or using a compressor It may be used to compress the mixed gas, or to use the cooler and the compressor at the same time.
- the above step can be performed, it may be performed by a compressor without limitation in its configuration and structure.
- the above step may be repeated until a high concentration of branched chain amino acid concentrate can be obtained.
- the step may be performed 2 to 50 times, 2 to 45 times, 2 to 40 times, 2 to 35 times, 2 to 30 times, 2 to 25 times, 2 to 20 times, 2 to 15 times, 2 to 10 times. Times, or may be repeated 2 to 5 times, but is not limited thereto.
- the method of the present application includes the step of reusing ammonia derived from the mixed gas obtained in step (c) as the ammonia of step (a), so that it does not require a large amount of pH adjusting material and is economical and branched-chain amino acid. It can improve the crystallization efficiency and can be used in an environmentally friendly way by reducing the generation of salt waste that is difficult to sustainably circulate.
- the method of the present application may further include, between steps (a) and (b), filtering the solution in which the branched chain amino acid crystals are dissolved, thereby removing the biomass in the solution. have.
- the method of the present application may further include, between steps (a) and (b), adding activated carbon to the solution in which the branched chain amino acid crystals are dissolved to remove the colored substances in the solution.
- ammonia generated in a plurality of crystallization processes could be obtained with a high level of recovery, and the recovered ammonia, for example, aqueous ammonia, could be reused to dissolve the branched chain amino acid crystals present in the reaction solution.
- the method can continuously produce branched-chain amino acid crystals based on the crystallization process without a separate pH adjusting agent, and thus can improve the production efficiency of branched-chain amino acid crystals.
- the method of the present application may further include, after step (b), separating the branched chain amino acid crystals from the concentrate containing the branched chain amino acid crystals of the step (b).
- Separating the branched chain amino acid crystal may be performed, for example, between steps (b) and (c), between steps (c) and (d), or after step (d). And, it may be made simultaneously with step (c) or step (d), but if it is after step (b), it may be made at any step regardless of the order of steps (c) and (d).
- the separation of the branched chain amino acid crystals from the concentrate includes, for example, solid-liquid separation of the concentrate containing the branched chain amino acid crystals to obtain a wet crystal of branched chain amino acids; and drying the obtained wet crystal to obtain branched chain amino acid crystals, but related to the separation and purification of amino acid crystals, conventional techniques known in the art may be applied without limitation.
- the present application is another aspect, (a) mixing the reaction solution containing the branched chain amino acid crystals with ammonia to obtain a solution in which the branched chain amino acid crystals are dissolved; (b) crystallizing the obtained solution to obtain a concentrated solution containing branched chain amino acid crystals; (c) obtaining a mixed gas including water vapor and ammonia, generated in the crystallization process; And (d) reusing the ammonia derived from the obtained mixed gas as the ammonia of the step (a); including, wherein the steps (b) and (c) are carried out simultaneously or sequentially, branched chain Branched chain amino acid crystals produced by a method for crystallizing amino acids are provided.
- the crystallization method of the branched chain amino acid is as described above.
- the method for crystallizing branched-chain amino acids according to the present application obtains ammonia, which was first added to increase the solubility of branched-chain amino acids in a vapor state during the crystallization process, and reuses it, thereby improving the production efficiency of branched-chain amino acid crystals, Production costs can be reduced because a large amount of pH adjusting material and additional neutralization process are not required.
- the method for crystallizing branched chain amino acids according to the present application can be utilized as an environmentally friendly method by further reducing the generation of salt wastes that are difficult to sustain sustainable circulation.
- L-valine, L-isoleucine, and L-leucine products with a purity of 98% or more were used, and CJ CheilJedang products were used as the branched chain amino acid products.
- water tertiary distilled water prepared by our was used, and 26% (v/v) ammonia water and 98% (v/v) sulfuric acid were purchased from Daejung Hwageum and used.
- 0.1M aqueous nitric acid solution and 0.02M aqueous dipicolinic acid solution for high-performance liquid chromatography (HPLC) analysis were purchased from Sigma-Aldrich (US) and used.
- the concentration of the branched-chain amino acid solution and the purity of the branched-chain amino acid crystals were analyzed using high-performance liquid chromatography (model DIONEX Ultimate 3000 system, Thermo Scientific, US), and the analysis conditions were as follows:
- ammonia concentration in the branched-chain amino acid solution was performed using ion chromatography (model 930 compact IC Flex, Metrohm, Switzerland), and the analysis conditions were as follows:
- the solubility of branched-chain amino acids was measured in a 1 L jacketed reactor made of glass. After mixing distilled water and ammonia water in various ratios in a 1 L jacketed reactor, an excess of branched-chain amino acid crystals was added and stirred, thereby preparing a reaction solution containing branched-chain amino acid crystals. Thereafter, while the internal temperature of the reactor was kept constant at 30° C. using a refrigeration/heating circulation device (model F35, Julabo, Germany), this condition was maintained for at least 12 hours with stirring.
- Crystallization was performed a total of 5 times under the same process conditions for each condition.
- ammonia water was added to a solution containing BCAA crystals prepared by mixing BCAA crystals and distilled water to dissolve the entire BCAA crystals.
- the recovered ammonia water was added to the reaction solution containing BCAA crystals to dissolve BCAA crystals (experimental group).
- a control group a group in which 26% (v/v) aqueous ammonia was added to a reaction solution containing BCAA crystals without a recovery process of ammonia water, and then the pH was readjusted to 7 with 98% (v/v) sulfuric acid. was used.
- Crystallization and ammonia recovery is a crystallizer comprising an injection unit, a pH control unit, a heating circulator and an outlet into which the crystallization feed is injected; a vapor recovery device including an injection unit into which the recovered ammonia water is injected, a cooling circulator and an outlet; And a device having a structure including a compressor between the vapor recovery device in the crystallizer was used.
- the crystallization feed refers to a concentrate containing branched chain amino acid crystals obtained in the first crystallization process, and the crystallization was carried out in a glass material 20 L jacketed reactor, and to prevent scaling in the heat exchange unit, the jacket The silver was installed only up to the height of the point where the volume of the internal liquid was 10 L.
- the jacket area was filled with tertiary distilled water whose temperature was controlled by a refrigeration/heating circulation device (model F35, Julabo, Germany), and the temperature was maintained at 80° C. during the crystallization process.
- Agitation of the internal reaction solution was carried out using a stirrer (model RW-20, IKA, Germany) capable of controlling the rotation speed with a 4-blade impeller made of Teflon, and the stirring speed was maintained at 200 rpm while crystallization was in progress.
- the pressure reduction inside the reactor was carried out using a compressor connected by a pipe, and an electronic vacuum controller (model NVC 2300-A, Eyela, Japan) was installed between the compressor and the crystallizer to control the pressure. adjusted.
- the pressure inside the crystallizer was maintained at 100 mbar.
- the ammonia recovery unit consisted of a 20 L jacketed pressure vessel made of stainless steel that can store up to 20 bar, and was cooled with cooling water at 4°C supplied to its own facility. At this time, the recovery of ammonia was carried out until the pH inside the crystallizer became 5.5 to 8.0.
- the concentration of the final concentrate was adjusted to be 1.5 times the concentration of the reaction solution containing BCAA crystals before ammonia was added.
- the crystallization process is terminated and the concentrate in the reactor is recovered through a discharge pipe disposed below. Thereafter, the recovered concentrate was subjected to solid-liquid separation for 5 min at a speed of 2000 rpm using a centrifugal basket separator (Model H-122, Kokusan, Japan) equipped with a cotton filter. If necessary, a washing process was performed at the beginning of the separation using distilled water. Thereafter, the obtained wet crystal was dried in an oven dryer at 80° C. until there was no change in weight to obtain BCAA crystals.
- Corynebacterium glutamicum a strain that produces L-leucine, a mutant (accession number: KCCM11662P), a strain that produces L-isoleucine, a Corynebacterium glutamicum mutant (accession number: KCCM11248P) , and a Corynebacterium glutamicum mutant strain (Accession No.: KCCM11336P), which is a strain producing L-valine, was used to prepare a fermentation broth containing L-leucine, L-isoleucine and L-valine, respectively.
- the pre-culture medium was dispensed into a 500 mL shaking Erlenmeyer flask, autoclaved at 121°C for 15 minutes, inoculated with each strain, and cultured for 24 hours in a rotary stirrer while stirring at 33°C at 200 rpm. did.
- the 5 L fermenter was filled with 3 L of the seed culture medium and autoclaved at 121° C. for 30 minutes, the pH was adjusted to 7.0, and 4% of the pre-culture was inoculated with 800 rpm at 33° C. and an aeration volume of 0.5 vvm.
- the seed culture was performed by culturing until the OD value reached 20.
- Seed culture medium Main culture medium Glucose (g/l) 5.0 10.1 40.2 MgSO 4 (g/l) 0.5 0.5 4.2 Yeast Extract (g/l) 5 10 3.2 KH 2 PO 4 2 3 3 Ammonium Sulfate (g/l) 6.3 NH 4 Cl (g/l) 0.5 One NaCl (g/l) 0.5 0.5 Na 2 HPO 4 (g/l) 4.07 5.07
- biomass such as cells was removed from the solution in which BCAA crystals were dissolved using a microfiltration device equipped with a 0.1 ⁇ m microfiltration membrane (model Pellicon 2, Merck, US), and for removal of colored substances, the After adding 10 wt% of powdered activated carbon (model YL303, Yuanli, China) compared to BCAA to the permeate, the mixture was stirred at 60° C. for 30 minutes. Thereafter, activated carbon was removed from the BCAA solution through primary vacuum filtration using a 7 ⁇ m filtration membrane, and residual activated carbon was further removed through secondary vacuum filtration using a 0.45 ⁇ m filtration membrane.
- powdered activated carbon model YL303, Yuanli, China
- L-leucine purification was repeated 5 times from the fermentation broth having an L-leucine concentration of 60 g/L containing L-leucine crystals prepared in (1) of Example 3 above.
- the pH was adjusted to 10 by adding the recovered ammonia water (however, the aqueous ammonia reagent) to 20 L of the fermentation broth containing L-leucine crystals, and after dissolving the entire amount of L-leucine, the biomass was removed through microfiltration. . Thereafter, the microfiltration permeate was treated with powdered activated carbon to separate colored substances. Thereafter, concentrated crystallization of the filtrate was carried out to a concentration of 90 g/L, and the concentrate containing L-leucine crystals was separated into solid and liquid using a basket filter. At this time, washing was performed using 20 vol% of tertiary distilled water based on the concentrate containing L- leucine crystals.
- the weight of the finally recovered crystals through drying was 1.1 kg on average 5 times, and the purity of the crystals was 98.4% on average 5 times.
- L-isoleucine Purification of L-isoleucine was repeated 5 times from the fermentation broth having an L-isoleucine concentration of 90 g/L containing L-isoleucine crystals prepared in (1) of Example 3 above.
- the recovered ammonia water (however, at first, ammonia water reagent) is added to adjust the pH to 10, the entire amount of L-isoleucine is dissolved, and the biomass is filtered through microfiltration. removed. Thereafter, the microfiltration permeate was treated with activated carbon powder to separate colored substances.
- the weight of the crystals finally recovered through drying was 1.7 kg on average 5 times, and the purity of the crystals was 98.6% on average 5 times.
- L-valine purification was repeated 5 times from the fermentation broth having an L-valine concentration of 150 g/L containing L-valine crystals prepared in (1) of Example 3 above.
- the recovered ammonia water (however, initially, aqueous ammonia reagent) was added to adjust the pH to 10, and after dissolving the entire amount of L-valine, the biomass was removed through microfiltration. Thereafter, the microfiltration permeate was treated with powdered activated carbon to separate colored substances. Thereafter, concentrated crystallization of the filtrate was carried out to a concentration of 225 g/L, and the concentrate containing L-valine crystals was separated into solid and liquid using a basket filter. At this time, washing was performed using 20 vol% of tertiary distilled water based on the concentrate containing L-valine crystals.
- the weight of the finally recovered crystals through drying was 3.0 kg on average 5 times, and the purity of the crystals was 98.4% on average 5 times.
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Abstract
Description
항목 | 대조군 | 실험군 | ||||
초기 반응액 농도 (g/L) | 30 | 100 | 200 | 30 | 100 | 200 |
초기 반응액 pH | 6 | 6 | 6 | 6 | 6 | 6 |
회수 암모니아 투입비 (vol%) | 0 | 0 | 0 | 4 | 8 | 13 |
26% 암모니아 투입비 (vol%) | 1 | 5 | 10 | 0 | 0 | 0 |
98% 황산 투입비 (vol%) | 1 | 2 | 4 | 1 | 2 | 4 |
1회차 결정화 과정에서 수득된 농축액의 농도 (g/L) | 29 | 93 | 175 | 28 | 91 | 167 |
1회차 결정화 과정에서 수득된 농축액의 pH | 7 | 7 | 7 | 9 | 11 | 12 |
최종 농축액 농도 (g/L) | 45 | 150 | 300 | 45 | 150 | 300 |
최종 농축액 pH | 7 | 7 | 7 | 7 | 7 | 7 |
결정 회수율 (%) | 42 | 81 | 90 | 42 | 81 | 90 |
암모니아 회수율 (%) | 0 | 0 | 0 | 97 | 98 | 98 |
회수 암모니아 농도 (%) | 0 | 0 | 0 | 9 | 17 | 23 |
항목 | 대조군 | 실험군 | ||||
초기 반응액 농도 (g/L) | 50 | 150 | 250 | 50 | 150 | 250 |
초기 반응액 pH | 6 | 6 | 6 | 6 | 6 | 6 |
회수 암모니아 투입비 (vol%) | 0 | 0 | 0 | 6 | 11 | 17 |
26% 암모니아 투입비 (vol%) | 2 | 7 | 12 | 0 | 0 | 0 |
98% 황산 투입비 (vol%) | 1 | 3 | 5 | 1 | 3 | 5 |
1회차 결정화 과정에서 수득된 농축액의 농도 (g/L) | 48 | 136 | 213 | 45 | 131 | 205 |
1회차 결정화 과정에서 수득된 농축액의 pH | 7 | 7 | 7 | 9 | 11 | 12 |
최종 농축액 농도 (g/L) | 75 | 225 | 375 | 75 | 225 | 375 |
최종 농축액 pH | 7 | 7 | 7 | 7 | 7 | 7 |
결정 회수율 (%) | 45 | 80 | 88 | 45 | 80 | 88 |
암모니아 회수율 (%) | 0 | 0 | 0 | 97 | 98 | 98 |
회수 암모니아 농도 (%) | 0 | 0 | 0 | 9 | 17 | 22 |
항목 | 대조군 | 실험군 | ||||
초기 반응액 농도 (g/L) | 120 | 250 | 350 | 120 | 250 | 350 |
초기 반응액 pH | 6 | 6 | 6 | 6 | 6 | 6 |
회수 암모니아 투입비 (vol%) | 0 | 0 | 0 | 11 | 21 | 30 |
26% 암모니아 투입비 (vol%) | 7 | 14 | 20 | 0 | 0 | 0 |
98% 황산 투입비 (vol%) | 3 | 6 | 8 | 1 | 3 | 5 |
1회차 결정화 과정에서 수득된 농축액의 농도 (g/L) | 110 | 209 | 275 | 107 | 206 | 273 |
1회차 결정화 과정에서 수득된 농축액의 pH | 7 | 7 | 7 | 9 | 11 | 12 |
최종 농축액 농도 (g/L) | 180 | 375 | 525 | 180 | 375 | 525 |
최종 농축액 pH | 7 | 7 | 7 | 7 | 7 | 7 |
결정 회수율 (%) | 76 | 87 | 90 | 76 | 87 | 90 |
암모니아 회수율 (%) | 0 | 0 | 0 | 98 | 98 | 98 |
회수 암모니아 농도 (%) | 0 | 0 | 0 | 13 | 17 | 23 |
조성 | 전배양 배지 | 종배양 배지 | 본배양 배지 |
포도당 (g/l) | 5.0 | 10.1 | 40.2 |
MgSO4 (g/l) | 0.5 | 0.5 | 4.2 |
효모추출액 (g/l) | 5 | 10 | 3.2 |
KH2PO4 | 2 | 3 | 3 |
암모늄 설페이트 (g/l) | 6.3 | ||
NH4Cl (g/l) | 0.5 | 1 | |
NaCl (g/l) | 0.5 | 0.5 | |
Na2HPO4 (g/l) | 4.07 | 5.07 |
Claims (12)
- (a) 분지쇄 아미노산 결정을 포함하는 반응액과 암모니아를 혼합하여 상기 분지쇄 아미노산 결정이 용해된 용해액을 수득하는 단계;(b) 상기 수득된 용해액을 결정화하여 분지쇄 아미노산 결정을 포함하는 농축액을 수득하는 단계;(c) 상기 결정화 과정에서 생성된, 수증기 및 암모니아를 포함하는 혼합 기체를 수득하는 단계; 및(d) 상기 수득한 혼합 기체로부터 유래한 암모니아를 상기 (a) 단계의 암모니아로 재사용하는 단계;를 포함하고,상기 (b) 단계 및 (c) 단계는 동시 또는 순차적으로 실시되는, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 분지쇄 아미노산은 L-루이신, L-이소루이신, 및 L-발린으로 이루어진 군에서 선택되는 적어도 하나의 아미노산인 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 분지쇄 아미노산 결정을 포함하는 반응액은 분지쇄 아미노산 결정을 포함하는 용액 또는 발효액을 포함하는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 3에 있어서, 상기 분지쇄 아미노산 결정을 포함하는 발효액은 분지쇄 아미노산을 생산하는 미생물을 배지에서 배양하여 수득되는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (a) 단계에서, 상기 분지쇄 아미노산 결정이 용해된 용해액의 pH는 9 내지 12인 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (b) 단계의 분지쇄 아미노산 결정을 포함하는 농축액의 분지쇄 아미노산 농도는 (a) 단계의 분지쇄 아미노산 결정을 포함하는 반응액의 분지쇄 아미노산 농도 대비 1.3 내지 1.7 배인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (c) 단계는 상기 (b) 단계의 분지쇄 아미노산 용해액의 pH가 5.5 내지 8.0이 될 때까지 실시하는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (d) 단계는 상기 암모니아를 증기 상태로 재사용하거나, 상기 증기가 압축 또는 응축된 액체 상태로 재사용하는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (a) 단계와 (b) 단계 사이에,(a-1) 상기 분지쇄 아미노산 결정이 용해된 용해액을 미세 여과하여, 용해액 내 바이오매스를 제거하는 단계;를 추가로 포함하는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (a) 단계와 (b) 단계 사이에,(a-2) 상기 분지쇄 아미노산 결정이 용해된 용해액에 활성탄을 첨가하여, 용해액 내 유색 물질을 제거하는 단계;를 추가로 포함하는 것인, 분지쇄 아미노산의 결정화 방법.
- 청구항 1에 있어서, 상기 (b) 단계 이후에,(e) 상기 (b) 단계의 분지쇄 아미노산 결정을 포함하는 농축액으로부터 분지쇄 아미노산 결정을 분리하는 단계;를 추가로 포함하는 것인, 분지쇄 아미노산의 결정화 방법.
- (a) 분지쇄 아미노산 결정을 포함하는 반응액과 암모니아를 혼합하여 상기 분지쇄 아미노산 결정이 용해된 용해액을 수득하는 단계;(b) 상기 수득된 용해액을 결정화하여 분지쇄 아미노산 결정을 포함하는 농축액을 수득하는 단계;(c) 상기 결정화 과정에서 생성된, 수증기 및 암모니아를 포함하는 혼합 기체를 수득하는 단계; 및(d) 상기 수득한 혼합 기체로부터 유래한 암모니아를 상기 (a) 단계의 암모니아로 재사용하는 단계;를 포함하고,상기 (b) 단계 및 (c) 단계는 동시 또는 순차적으로 실시되는, 분지쇄 아미노산의 결정화 방법에 의해 생산된 분지쇄 아미노산 결정.
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JP2022528572A JP7350174B2 (ja) | 2019-11-25 | 2020-11-09 | アンモニアの持続可能な循環のできる分枝鎖アミノ酸の結晶化方法 |
CN202080073444.1A CN114599634B (zh) | 2019-11-25 | 2020-11-09 | 允许氨的循环能够持续的支链氨基酸的结晶化方法 |
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