Method for producing high-purity arginine

Abstract

The present application relates to a method for producing arginine, wherein a bleaching step is not performed, and an anion exchange step is separated and performed simultaneously with an arginine production step, thereby improving the yield of arginine and significantly reducing the time required for production.

Description

Method for producing high-purity arginine

[0001] Cross-citation with related application(s)

[0002] This application claims the benefit of priority from Republic of Korea Patent Application No. 10-2023-0117455, dated September 5, 2023, the entire contents of which are incorporated herein by reference.

[0003] The present application relates to a method for producing high-purity arginine.

[0004]

[0005] The present invention relates to a method for producing high-purity arginine used in animal feed additives, etc., from an arginine fermentation solution. Existing arginine production methods include separation of cells from an arginine fermentation solution, a cation exchange process, a decolorization process, an anion exchange process, and then crystallization and separation steps. Among these, the decolorization process is a step in which activated carbon is introduced to enable the adsorption and separation of organic substances or colored ionic substances contained in the arginine process solution, thereby improving the purity of the arginine process solution and the light transmittance of arginine. However, recently, the demand for light transmittance of arginine products used in animal feed additives, etc., has been decreasing, and environmental issues such as waste activated carbon and wastewater treatment generated during the decolorization process are being highlighted.

[0006]

[0007] [Prior Art Literature]

[0008] (Patent Document 1) U.S. Patent Publication No. US 2003-0124686 A1

[0009]

[0010] One purpose of the present application is to provide a method for producing arginine.

[0011]

[0012] The present application relates to a method for manufacturing arginine, in which the yield of arginine is improved and the time required for manufacturing arginine is drastically reduced by performing an anion exchange process separately and simultaneously with the arginine manufacturing process without performing a decolorization process.

[0013]

[0014] One aspect provides a method for producing arginine.

[0015] The above arginine may be arginine crystals or arginine powder.

[0016]

[0017] Glossary of terms

[0018] In this application, the term "fermented product" may refer to the result of enzymatic or metabolic decomposition of an organic substance using a microorganism. For example, the fermented product may include a culture obtained by culturing a microorganism in a culture medium, a culture obtained by removing the cells (the microorganism or a part of the microorganism) from the culture, or a concentrate, dried product, or lyophilized product of the culture from which the cells have been removed. Furthermore, in this case, the term "fermented liquid" may include the entire fermented product containing arginine produced by the microorganism, or a fermented product from which impurities have been removed.

[0019] In this application, the term “eluent” may mean a liquid that has passed through a cation exchange resin and / or an anion exchange resin.

[0020] In this application, the term "process liquid" may collectively refer to a liquid introduced into each process for manufacturing arginine (e.g., arginine crystals) and / or a liquid obtained after each process is performed. In one example, the process liquid may refer to a fermentation liquid, eluate, concentrate, filtrate, etc. containing arginine.

[0021] In the present application, the term "mother liquid" may mean a liquid remaining after performing a concentration crystallization process on a process liquid containing arginine to separate arginine (e.g., arginine crystals).

[0022]

[0023] Fermented liquid containing arginine

[0024] One aspect may provide a method for producing arginine crystals from a fermentation solution containing arginine.

[0025] The fermented solution containing the above arginine can be obtained by culturing a microorganism that produces arginine. The microorganism that produces arginine includes both wild-type microorganisms and microorganisms that have undergone natural or artificial genetic modification. It may be a microorganism in which a specific mechanism has been weakened or strengthened due to reasons such as the insertion of an external gene or the enhanced or inactivated activity of an endogenous gene, and may be a microorganism that includes a genetic modification of a target protein for arginine production.

[0026]

[0027] The arginine-producing microorganism of the present application may be, but is not limited to, a microorganism that naturally has arginine-producing ability, or a microorganism that has been granted arginine-producing ability to a microorganism that does not have arginine-producing ability. Specifically, the arginine-producing microorganism of the present application, or the microorganism having arginine-producing ability, may be a microorganism in which some of the genes in the arginine biosynthetic pathway are strengthened or weakened, or some of the genes in the arginine degradation pathway are strengthened or weakened. The phrase “strengthening” or “increasing” the arginine-producing ability of the microorganism of the present application means that the arginine-producing ability of the microorganism of the present application is improved compared to other microorganisms other than the microorganism of the present application, a parent strain, or an unmodified microorganism. For example, the microorganism of the present application may have an arginine productivity that is improved by about 1% or more, 2% or more, 5% or more, 10% or more, 100% or more, 200% or more, 500% or more, 1000% or more, 1100% or more, 1200% or more, or 1300% or more compared to the arginine productivity of other microorganisms, but is not limited thereto. The term “about” includes all of ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc., and includes all of the values ​​in a range equal to or similar to the value following the term “about,” but is not limited thereto.

[0028] The above arginine-producing microorganism may be an Escherichia coli microorganism, a Bacillus sp. microorganism, a Saccharomyces sp. microorganism, or a Corynebacterium sp. If the above microorganism is an Escherichia coli microorganism, the microorganism may be Escherichia coli. If the above microorganism is a Bacillus microorganism, the microorganism may be at least one selected from the group consisting of Bacillus subtilis, Bacillus vulgaris, Bacillus cereus, etc. If the above microorganism is a Saccharomyces microorganism, the microorganism may be Saccharomyces cerevisiae.If the above microorganism is a microorganism of the genus Corynebacterium, the microorganism is Corynebacterium glutamicum, Corynebacterium erythrogenes, Corynebacterium crudilactis, Corynebacterium deserti, Corynebacterium efficiens, Corynebacterium callunae, Corynebacterium stationis, Corynebacterium singulare, Corynebacterium halotolerans, Corynebacterium striatum, It may be at least one selected from the group consisting of Corynebacterium ammoniagenes, Corynebacterium pollutisoli, Corynebacterium imitans, Corynebacterium testudinoris, Corynebacterium crenatum, and Corynebacterium flavescens, and may be, for example, Corynebacterium glutamicum, but is not limited thereto.

[0029] The cultivation of the above arginine-producing microorganism can be carried out according to an appropriate medium and culture conditions known in the art. This culture process can be easily adjusted and used by a person skilled in the art according to the selected strain. Specifically, the culture may be batch, continuous, or fed-batch, but is not limited thereto. The term "medium" in this application refers to a material mixed with nutrients as the main component necessary for culturing the microorganism, and supplies nutrients and growth factors, including water, which are essential for survival and growth. Specifically, the medium and other culture conditions used for culturing the microorganism of this application can be any medium used for culturing general microorganisms without particular limitation. However, the microorganism of this application can be cultured under aerobic conditions by controlling temperature, pH, etc. in a general medium containing an appropriate carbon source, nitrogen source, phosphorus source, inorganic compound, amino acid, and / or vitamin, etc.

[0030] In one example, the fermentation solution containing the arginine may be one from which the microorganisms (e.g., microorganisms producing arginine, or parts of the microorganisms, etc.) have been removed.

[0031] Removal of the fungal cells from the above fermentation solution may be accomplished by conventional methods such as separation (e.g., centrifugation, etc.) or filtration (e.g., use of a fungal cell separation facility, ceramic membrane filter, etc.).

[0032]

[0033] (a) Step: Passing the fermentation solution containing arginine through a cation exchange resin (cation exchange process)

[0034] The method for producing arginine of the present application may include a step of passing a fermentation solution containing the arginine through a cation exchange resin.

[0035] The cation exchange resin can remove cationic impurities. The cation exchange resin can be a commonly known cation exchange resin, such as a strongly acidic cation exchange resin (SAC) or a weakly acidic cation exchange resin (WAC), but is not limited thereto. The cation exchange resin can be a gel type or a porous type, but is not limited thereto.

[0036] The cationic impurity may be an alkali metal ion (e.g., sodium ion, potassium ion, etc.), an alkaline earth metal ion (e.g., magnesium ion, calcium ion, etc.), or an ammonium ion.

[0037] The above cation exchange process may include a step of treating a eluent to desorb arginine adsorbed to the cation exchange resin.

[0038] The above eluent may be any substance commonly used to separate a substance remaining on a stationary phase without limitation. In one example, sodium hydroxide or an aqueous ammonia solution may be used as the eluent, but is not limited thereto.

[0039] In one example, the eluent can be used by diluting it in various concentrations, and in one example, 0.01 to 5N, 0.01 to 4N, 0.01 to 3N, 0.01 to 2N, 0.01 to 1N, 0.05 to 5N, 0.05 to 4N, 0.05 to 3N, 0.05 to 2N, 0.05 to 1N, 0.1 to 5N, 0.1 to 4N, 0.1 to 3N, 0.1 to 2N, 0.1 to 1N, 0.5 to 5N, 0.5 to 4N, 0.5 to 3N, 0.5 to 2N, 0.5 to 1N, 1 to 5N, 1 to 4N, 1 to 3N, 1 to 2N, 2 to It can be used in concentrations of 5N, 2 to 4N, 2 to 3N, 2.5 to 5N, 2.5 to 4N, or 2.5 to 3N.

[0040] The above cation exchange process may include a step of concentrating under high temperature conditions to remove the used eluent from the eluent obtained in the cation exchange process after the step of treating the eluent.

[0041] The above high temperature condition can be set without limitation as long as it is a condition that can remove the used eluent (e.g., aqueous ammonia solution, etc.), and in one example, it can be 30°C or higher, 35°C or higher, 40°C or higher, 45°C or higher, 50°C or higher, 55°C or higher, or 60°C or higher.

[0042] The above-mentioned concentrating step may be a step of concentrating to a concentration before arginine is obtained as a crystal.

[0043] The above-mentioned concentrating step is performed such that the concentration of arginine in the concentrate is 100 g / l or less, 110 g / l or less, 120 g / l or less, 130 g / l or less, 140 g / l or less, 150 g / l or less, 160 g / l or less, 170 g / l or less, 180 g / l or less, 190 g / l or less, 200 g / l or less, 210 g / l or less, 220 g / l or less, 230 g / l or less, 240 g / l or less, 250 g / l or less, 30 to 100 g / l, 30 to 110 g / l, 30 to 120 g / l, 30 to 130 g / l, 30 to 140 g / l, 30 to 150 g / l, 30 to 160 g / l, 30 to 170 g / l, 30 to 180 g / l, 30 to 190 g / l, 30 to 200 g / l, 30 to 210 g / l, 30 to 220 g / l, 30 to 230 g / l, 30 to 240 g / l, 30 to 250 g / l, 40 to 100 g / l, 40 to 110 g / l, 40 to 120 g / l, 40 to 130 g / l, 40 to 140 g / l, 40 to 150 g / l, 40 to 160 g / l, 40 to 170 g / l, 40 to 180 g / l, 40 to 190 g / l, 40 to 200 g / l, 40 to 210 g / l, 40 to 220 g / l, 40 to 230 g / l, 40 to 240 g / l, 40 to 250 g / l, 50 to 100 g / l, 50 to 110 g / l, 50 to 120 g / l, 50 to 130 g / l, 50 to 140 g / l, 50 to 150 g / l, 50 to 160 g / l, 50 to 170 g / l, 50 to 180 g / l, 50 to 190 g / l, 50 to 200 g / l, 50 to 210 g / l, 50 to 220 g / l, 50 to 230 g / l, 50 to 240 g / l, 50 to 250 g / l, 60 to 100 g / l, 60 to 110 g / l, 60 to 120 g / l, 60 to 130 g / l,60 to 140 g / l, 60 to 150 g / l, 60 to 160 g / l, 60 to 170 g / l, 60 to 180 g / l, 60 to 190 g / l, 60 to 200 g / l, 60 to 210 g / l, 60 to 220 g / l, 60 to 230 g / l, 60 to 240 g / l, 60 to 250 g / l, 70 to 100 g / l, 70 to 110 g / l, 70 to 120 g / l, 70 to 130 g / l, 70 to 140 g / l, 70 to 150 g / l, 70 to 160 g / l, 70 to 170 g / l, 70 It may be a step of concentrating to 180 g / l, 70 to 190 g / l, 70 to 200 g / l, 70 to 210 g / l, 70 to 220 g / l, 70 to 230 g / l, 70 to 240 g / l, or 70 to 250 g / l, but is not limited thereto.

[0044]

[0045] (b) Step: Step of concentrating and crystallizing the process solution obtained in step (a) (concentration and crystallization process)

[0046] The method for producing arginine of the present application may include a step of concentrating and crystallizing the process solution obtained in step (a) (concentration and crystallization step).

[0047] The process solution obtained in the above step (a) may be an eluent obtained by subjecting the fermentation solution containing arginine to a cation exchange process or a concentrate obtained by subjecting the eluent to a concentration process.

[0048] The above concentration crystallization process may be a step of concentrating arginine to a concentration at which it is obtained as a crystal. The above concentration crystallization process is such that the concentration of arginine in the concentrate is 200 g / l or more, 250 g / l or more, 300 g / l or more, 350 g / l or more, 400 g / l or more, 450 g / l or more, 500 g / l or more, 550 g / l or more, 600 g / l or more, 650 g / l or more (the upper limit is about 700 g / l), 200 to 700 g / l, 200 to 650 g / l, 200 to 600 g / l, 200 to 550 g / l, 200 to 500 g / l, 250 to 700 g / l, 250 to 650 g / l, 250 to 600 g / l, 250 to 550 g / l, 250 to 500 g / l, 300 to 700 g / l, 300 to 650 g / l, 300 to 600 g / l, 300 to 550 g / l, 300 to 500 g / l, 350 to 700 g / l, 350 to 650 g / l, 350 to 600 g / l, 350 to 550 g / l, 350 to 500 g / l, 400 to 700 g / l, 400 to 650 g / l, 400 to 600 g / l, 400 to 550 g / l, 400 to 500 g / l, 450 to 700 g / l, 450 to 650 g / l, 450 to 600 g / l, 450 It may be a step of concentrating to, but is not limited to, 550 g / l, 450 to 500 g / l, 500 to 700 g / l, 500 to 650 g / l, 500 to 600 g / l, or 500 to 550 g / l.

[0049] The above concentration crystallization process can be performed under temperature conditions that can obtain arginine as crystals. In one example, the concentration crystallization process is performed at a temperature of 30 to 80°C, 30 to 75°C, 30 to 70°C, 30 to 65°C, 30 to 60°C, 30 to 55°C, 30 to 50°C, 35 to 80°C, 35 to 75°C, 35 to 70°C, 35 to 65°C, 35 to 60°C, 35 to 55°C, 35 to 50°C, 40 to 80°C, 40 to 75°C, 40 to 70°C, 40 to 65°C, 40 to 60°C, 40 to 55°C, 40 to 50°C, 45 to 80°C, 45 to 75°C, 45 to 70°C, 45 to 65°C, 45 to It can be performed at 60°C, 45 to 55°C, 45 to 50°C, 50 to 80°C, 50 to 75°C, 50 to 70°C, 50 to 65°C, 50 to 60°C, 50 to 55°C, for example, but is not limited thereto.

[0050] The above concentration crystallization process may additionally include a cooling step to reduce the solubility of arginine and thereby increase the yield of arginine crystals. The above cooling step is to cool the arginine concentrate to 40 ℃ or less, 35 ℃ or less, 30 ℃ or less, 25 ℃ or less, 20 ℃ or less, 15 ℃ or less, 10 ℃ or less, 5 ℃ or less (the lower limit is 0 ℃), 0 to 40 ℃, 0 to 35 ℃, 0 to 30 ℃, 0 to 25 ℃, 0 to 20 ℃, 0 to 15 ℃, 0 to 10 ℃, 0 to 5 ℃, 5 to 40 ℃, 5 to 35 ℃, 5 to 30 ℃, 5 to 25 ℃, 5 to 20 ℃, 5 to 15 ℃, 5 to 10 ℃, 10 to 40 ℃, 10 to 35 ℃, 10 to 30 ℃, 10 to The step may be a cooling step at 25 ℃, 10 to 20 ℃, 10 to 15 ℃, 15 to 40 ℃, 15 to 35 ℃, 15 to 30 ℃, 15 to 25 ℃, 15 to 20 ℃, 20 to 40 ℃, 20 to 35 ℃, 20 to 30 ℃, 20 to 25 ℃, 25 to 40 ℃, 25 to 35 ℃, 25 to 30 ℃, but is not limited thereto.

[0051] The above cooling step can use a conventional cooling method, such as using a cooler.

[0052]

[0053] In the above concentration crystallization process, arginine can be obtained, and the remaining liquid after obtaining arginine can be separated as a mother liquor.

[0054] The above step (b) may additionally include a step of drying the obtained arginine, and by drying the moisture, high-purity arginine can be finally obtained.

[0055] The above arginine may be arginine crystals or arginine powder.

[0056]

[0057] The conventional arginine manufacturing method had problems with waste activated carbon and wastewater treatment generated during the decolorization process, including the decolorization process within the arginine manufacturing process, and with the anion exchange process within the arginine manufacturing process, which took a long time to manufacture arginine.

[0058] However, the arginine manufacturing method of the present application can achieve the effect of improving the yield of arginine and drastically reducing the time required for arginine manufacturing by separately separating the anion exchange process and performing it simultaneously with the arginine manufacturing process without performing the decolorization process.

[0059] Therefore, the arginine manufacturing method of the present application may be characterized by separating the mother liquor separated in the above-mentioned concentration crystallization step from the arginine manufacturing process, passing it through an anion exchange resin in a separate process to remove impurities, and then reusing the obtained eluent in step (a), step (b), or steps (a) and (b). This step will be described in more detail in step (c) below.

[0060]

[0061] (c) A step of passing the mother liquor obtained in step (b) through an anion exchange resin (anion exchange process)

[0062] The method for producing arginine of the present application may include a step of passing the mother liquor separated in the step (b) (concentration crystallization step) through an anion exchange resin.

[0063] The anion exchange resin can remove anionic impurities. The anion exchange resin can be a commonly known anion exchange resin, such as a strongly basic anion exchange resin (SBA) or a weakly basic anion exchange resin (WBA), but is not limited thereto. The anion exchange resin can be a gel type or a porous type, but is not limited thereto.

[0064] The above anionic impurity is sulfate (SO4 2- ), phosphate (PO4 2- ), chloride ion (Cl - ) but are not limited thereto.

[0065] The mother liquor that has passed through the above anion exchange resin can have anionic impurities removed, thereby improving the color of the liquid or the final product, arginine.

[0066] The above step (c) may be characterized in that it is performed spatially separated from the above steps (a) and (b).

[0067] That is, the anion exchange process of step (c) may be characterized in that it is spatially separated from the main process of producing arginine, i.e., the cation exchange process of step (a) and the concentration crystallization process of step (b), and is performed simultaneously.

[0068] The anion exchange process of step (c) above is a process that takes a long time, but by performing it simultaneously with steps (a) and (b) spatially separated from each other, the total time required for producing arginine can be drastically reduced.

[0069]

[0070] (d) a step of reusing the eluent obtained in step (c) above;

[0071] Arginine is a highly soluble amino acid, and since the mother liquor separated in step (b) also contains a large amount of arginine, it is important to effectively recycle the mother liquor. Accordingly, the arginine production method of the present application may include a step of mixing the eluent from the mother liquor that has undergone the anion exchange process with the arginine process solution from step (a), step (b), or steps (a) and (b) and repeating the process.

[0072] Specifically, the arginine process solution of step (a) may refer to (i) a fermentation solution containing arginine of step (a), (ii) an eluate obtained by subjecting the fermentation solution containing arginine to a cation exchange process, or (iii) a concentrate obtained by subjecting the eluate to a concentration process.

[0073] The arginine process solution of step (b) may be a liquid in which the concentration crystallization process of step (b) is performed, i.e., an eluent obtained by subjecting the fermentation solution containing arginine in step (a) to a cation exchange process or a concentrate obtained by subjecting the eluent to a concentration process.

[0074] In this way, the mother liquor separated in step (b) is spatially separated from the arginine manufacturing process, and the anion exchange process is performed as a separate process, and the eluent that has undergone the anion exchange process is mixed with the arginine process solution of step (a) and / or step (b) of the arginine manufacturing process, so that the anion exchange process and the arginine manufacturing process can be performed simultaneously, thereby drastically reducing the time required for arginine manufacturing.

[0075]

[0076] In the arginine manufacturing method of the present application, steps (a) to (d) may be characterized in that they are performed repeatedly.

[0077] That is, the arginine manufacturing method of the present application comprises the steps of (a): passing a fermentation solution containing arginine through a cation exchange resin (cation exchange process);

[0078] (b) Step: A step of concentrating and crystallizing the process solution obtained in step (a) (concentration and crystallization process);

[0079] (c) a step of passing the mother liquor obtained in step (b) through an anion exchange resin (anion exchange process); and

[0080] (d) a step of reusing the eluent obtained in step (c) in step (a), step (b), or steps (a) and (b),

[0081] The above steps (a) to (d) may be characterized in that they are performed repeatedly.

[0082]

[0083] The arginine manufacturing method of the present application may be characterized in that it does not perform a decolorization process.

[0084] The arginine manufacturing method of the present application does not require a decolorization process, yet exhibits no significant difference in the quality (purity, color, BD (Bulk Density)) of the final arginine produced. Therefore, processes such as waste activated carbon and wastewater treatment generated during the decolorization process are not required, thereby achieving environmentally friendly and economical benefits. Furthermore, by eliminating the decolorization process, the time required for the decolorization process can be reduced.

[0085] The arginine manufacturing method of the present application separates the anion exchange process and performs it simultaneously with the arginine manufacturing process, thereby improving the yield of arginine and drastically reducing the time required for arginine manufacturing compared to an arginine manufacturing method in which the anion exchange process is not separated separately.

[0086] In one example, the method for producing arginine of the present application can improve the yield of arginine by 1% or more, 2% or more, or 3% or more compared to an arginine producing method that includes a decolorization process and / or does not separately separate the anion exchange process, and due to this improvement in the yield of arginine, the amount of raw materials and auxiliary materials used in producing arginine can be reduced, thereby significantly reducing the manufacturing cost.

[0087] In one example, the method for producing arginine of the present application can reduce the time required for producing arginine by 1% or more, 2% or more, 3% or more, 4% or more, 5% or more, 6% or more, 7% or more, 8% or more, 9% or more, 10% or more, 15% or more, 20% or more, 25% or more, 30% or more, 35% or more, or 40% or more compared to a method for producing arginine that includes a decolorization process and / or does not include a separate anion exchange process.

[0088] In one example, the method for producing arginine of the present application can reduce the time required for producing arginine by about 1 hour or more, 2 hours or more, 3 hours or more, 4 hours or more, 5 hours or more, 6 hours or more, 7 hours or more, 8 hours or more, 9 hours or more, 10 hours or more, 15 hours or more, 20 hours or more, 25 hours or more, or 30 hours or more compared to a method for producing arginine that includes a decolorization process and / or does not include a separate anion exchange process.

[0089]

[0090] The present application relates to a method for manufacturing arginine in which the yield of arginine is improved and the time required for manufacturing is drastically reduced by performing an anion exchange process separately and simultaneously with the arginine manufacturing process without performing a decolorization process.

[0091]

[0092] Figure 1 shows a schematic diagram of an arginine manufacturing method including a conventional decolorization process and the manufacturing method of the present application.

[0093]

[0094] The present invention will be described in more detail below with reference to the following examples. However, these examples are provided solely to illustrate the present invention, and the scope of the present invention is not limited by these examples.

[0095]

[0096] The present application relates to a method for manufacturing arginine, wherein the yield of arginine is improved and the time required for arginine manufacturing is drastically reduced by performing a separate anion exchange process simultaneously with the arginine manufacturing process without performing a decolorization process. To verify the effectiveness of the present application, a comparison was made between the arginine manufacturing method of the present application and a conventional arginine manufacturing method that includes a decolorization process and performs the anion exchange process as part of the arginine manufacturing process, and the results are presented below.

[0097]

[0098] Comparative Example 1. Conventional arginine manufacturing method

[0099] a) Obtaining a fermentation solution containing arginine

[0100] To produce high-purity arginine using this process, arginine-producing microorganisms were cultured to obtain a fermentation solution (arginine fermentation solution) containing arginine having the composition shown in Table 1 below. The arginine fermentation solution includes cultured microorganisms and medium components, and the moisture content and composition were analyzed, as shown in Table 1 below.

[0101] Composition analysis of arginine fermentation solution Composition value: Arginine 130.6 g / L, amino acids other than arginine 2.6 g / L, minerals 62.0 g / L, organic acids 0.3 g / L, moisture 79.2%

[0102] The pH of the above arginine fermentation solution was adjusted downward to 4.0 by treating it with sulfuric acid, and the cells used for arginine fermentation were separated or filtered using a cell separation facility (membrane filter, mechanical separator, etc.).

[0103]

[0104] b) Step of passing the arginine fermentation solution through a cation exchange resin (cation exchange process)

[0105] The arginine fermentation solution from which the above-mentioned cells were removed was passed through a cation exchange resin (MC-08, Samyang Corporation) to obtain an arginine eluate in which anions were separated based on the adsorption / elution principle between the resin and arginine. The anion separation efficiency in the cation exchange process was approximately 99%. Ammonia (Daejung Chemicals & Metals) was used as an eluent to desorb the arginine adsorbed on the resin in the cation exchange process.

[0106]

[0107] c) Step of removing ammonia and concentrating arginine

[0108] The arginine eluent (liquid that passed through the ion exchange resin) obtained in step b) above was subjected to a concentration process at an internal temperature of 55°C using a post-stage vacuum Rotary Evaporator (BUCHI, Rotavapor R-300) to remove ammonia and increase the arginine concentration from approximately 72 g / L to 200 g / L.

[0109]

[0110] d) A step of passing the concentrate obtained in c) through an anion exchange resin (anion exchange process)

[0111] All of the concentrate obtained in step c) was passed through an anion exchange resin (AMP-26, Samyang Corporation) to adsorb and remove coexisting byproducts, and the arginine process solution was recovered.

[0112]

[0113] e) Activated carbon treatment step (decolorization process)

[0114] To improve the transmittance of the process solution passing through the anion exchange resin, the temperature of the process solution was raised to 60°C, activated carbon was added at a ratio of 3 wt% to the weight of arginine, and stirred for approximately 1 hour. Thereafter, the activated carbon was removed using a Nutsche filter and filter paper, and the filtrate was obtained.

[0115]

[0116] f) A step of performing a concentration crystallization process on the filtrate obtained in the above e) to obtain arginine crystals (concentration crystallization process)

[0117] The above filtrate is a high-purity arginine process solution, and the concentration crystallization process uses a vacuum rotary evaporator to concentrate the arginine concentration from 200 g / L to about 500-550 g / L at an internal temperature of 50°C, so that arginine exceeding the solubility is precipitated as crystals. After the concentration crystallization is complete, the solution is cooled to 25 to 30°C using a cooling circulator to increase the yield of arginine crystals by decreasing the solubility through lowering the temperature. The precipitated arginine crystals and the mother liquor are separated using a solid-liquid separator (such as a basket separator). The separated arginine crystals are dried to remove moisture through an oven dryer to ultimately obtain high-purity arginine.

[0118]

[0119] g) a step of reusing the mother liquor obtained in e) above;

[0120] Arginine is an amino acid with relatively high solubility, and the mother liquor generated during the separation process of arginine crystals (i.e., the liquid remaining after performing a concentration crystallization process on the process solution containing arginine to separate arginine crystals) also contains a large amount of arginine. Therefore, in order to increase the recovery rate of the process, the mother liquor was recovered as the arginine eluent of step b) and / or the arginine process solution of step f), and the process was repeated.

[0121]

[0122] The properties of each process solution of the existing arginine manufacturing method of Comparative Example 1 are summarized in Table 2 below.

[0123] Arginine Comparison of properties of process liquids of existing process methods Item Unit Process liquid Fermentation liquid Cell separation Cation resin Concentration Anion resin Decolorization / filtration Experimental equipment / materials - Membrane Filter Cation resin (MC-08) Rotary Evaporator Anion resin (AMP-26) Activated carbon Nut filter Retention time hr 45 10 12 20 6 pH - 7.0 4.0 1 1.6 10.7 10.7 1 1.1 Temperature ℃ 30 60 45 55 40 60 Concentration g / L 1 30 1 30 7 2 20 0 2 0 5 20 4 Purity % 5 6.1 6 0.0 9 3.1 9 3.4 9 6.1 9 6.6 * Color abs 1 4 1.6 12.10 4 3 1.1 7 0.0 3 0.0 2

[0124] * Color: Color Value at 430 nm (the lower the value, the clearer the liquid)* Purity: (Arginine weight / solid weight of process solution) * 100 As can be seen in Table 2 above, the anion exchange process is a step that takes a lot of time in the entire process.

[0125]

[0126] Example 1. Method for producing arginine without decolorization process

[0127] In the conventional arginine manufacturing method described in Comparative Example 1 above, (1) the decolorization process (step e)) was not performed, and (2) the anion exchange process (step d)) was performed separately and the mother liquor generated in step f) was processed, thereby performing the process simultaneously with the arginine manufacturing process.

[0128]

[0129] That is, steps a) to c) of Comparative Example 1 were performed in the same manner, and the arginine concentrate obtained in step c) was immediately subjected to the concentration crystallization step of step f), thereby separating the arginine crystals and the mother liquor.

[0130]

[0131] The anion exchange process is a time-consuming process, and in the present application, the anion exchange process is separated separately, and the mother liquor separated in the above-mentioned concentration crystallization step is subjected to a separate anion exchange process, and the obtained eluent (mother liquor passed through an anion exchange resin) is mixed with the arginine eluent of step b) and / or the concentrate to be input into the concentration crystallization process of step f), and the process is repeated.

[0132]

[0133] The quality of the mother liquor obtained in the arginine manufacturing process and the eluent obtained by passing it through an anion exchange resin are summarized in Table 3 below.

[0134] Process solution item unit mother liquor anion decolorization experiment average 1st time 2nd time 3rd time Mother liquor purity % 87.9 87.4 88.0 88.4 Chromaticity abs 1.75 1.64 1.84 1.76 Eluent purity % 92.4 93.0 92.89 1.5 Chromaticity abs 0.1 10.1 70.0 70.0 9 Purity difference % 4.5 5.7 4.8 3.2 Decolorization rate % 93.7 89.7 96.2 95.2

[0135] * Chromaticity: Color Value at 430nm (the lower the value, the clearer the liquid)

[0136] As shown in Table 3 above, the eluent that passed through the anion exchange resin had improved purity and color compared to the mother liquor.

[0137]

[0138] The above eluent was mixed with the arginine eluent of step b) (cation exchange process) and the concentrate to be introduced into step f (concentration crystallization process), and the process was repeated.

[0139]

[0140] 3. Compare results

[0141] When the quality of arginine manufactured by reusing the eluent obtained by passing the mother liquor separated during the arginine manufacturing process through an anion exchange resin without performing the decolorization process as in Example 1 was compared with that of arginine manufactured through the existing manufacturing method (Comparative Example 1), there was no significant difference in the main qualities (purity, pH, BD, etc.) (Table 4 below).

[0142]

[0143] Key quality and process indicators comparison items Detailed items Unit comparison Example 1 Example 1 Product quality Purity % 99.3 9 8.7 pH -10.8 10.7 Bulk Density g / ml 700 ± 20 680 ± 30 Process processing time Hours 7246 Process yield % 88.5 9 0.5

[0144] However, the arginine manufacturing method of the present application reduces the arginine decomposition loss that occurs during the decolorization process by not performing the decolorization process, thereby increasing the yield of the manufacturing process by about 2%, reducing the time required for the decolorization process, and by performing the anion exchange process separately and simultaneously from the arginine manufacturing process, reducing the time required for the anion exchange process, thereby achieving the effect of improving productivity through a time reduction of about 36% of the entire process.

[0145]

[0146] In addition, by not performing the decolorization process, the cost required for the decolorization process can be reduced, an environmentally friendly effect can be achieved as the treatment of waste activated carbon is unnecessary, and the yield of arginine is improved, thereby reducing the amount of raw materials and auxiliary materials, which can significantly reduce the manufacturing cost.

[0147]

[0148] From the above description, those skilled in the art will understand that the present invention can be implemented in other specific forms without altering its technical spirit or essential characteristics. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not restrictive. The scope of the present invention should be interpreted as encompassing all changes or modifications derived from the meaning and scope of the following claims and their equivalent concepts, rather than the detailed description above.

Claims

1. (a) A step of passing a fermentation solution containing arginine through a cation exchange resin to obtain an eluent; (b) a step of concentrating and crystallizing the eluent obtained in step (a); (c) a step of passing the mother liquor obtained in step (b) through an anion exchange resin to obtain an eluent; and (d) a step of reusing the eluent obtained in step (c) in step (a), step (b), or steps (a) and (b), The above steps (a) to (d) are characterized in that they are performed repeatedly. Method for producing arginine.

2. A method for producing arginine, wherein the fermentation liquid containing arginine in the first paragraph is a fermentation product obtained by culturing a microorganism producing arginine, or a fermentation product from which the microorganisms have been removed.

3. A method for producing arginine, wherein in the first paragraph, step (a) further includes a step of concentrating the eluent that has passed through the cation exchange resin to 200 g / l or less.

4. A method for producing arginine, wherein in the first paragraph, step (b) is to concentrate arginine to a concentration of 200 g / l or more.

5. A method for producing arginine, wherein in the first paragraph, step (b) is concentrated at 30 to 60°C.

6. A method for producing arginine, wherein step (b) further comprises a step of cooling the concentrate to 35°C or lower.

7. A method for producing arginine, wherein step (b) further comprises a step of drying the obtained arginine.

8. In paragraph 1, A method for producing arginine, characterized in that the step (c) is performed simultaneously with the steps (a) and (b).

9. A method for producing arginine, characterized in that no decolorization process is performed in any one of claims 1 to 8.

10. A method for producing arginine, wherein the time required for producing arginine is reduced by 20% or more compared to a method for producing arginine in which the anion exchange process is not separately separated according to any one of paragraphs 1 to 8.

11. A manufacturing method according to any one of claims 1 to 8, wherein the manufacturing method of arginine is a manufacturing method of arginine crystals or arginine powder.

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