CN112608254A - Method for preparing L-citrulline - Google Patents
Method for preparing L-citrulline Download PDFInfo
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- CN112608254A CN112608254A CN202011586600.2A CN202011586600A CN112608254A CN 112608254 A CN112608254 A CN 112608254A CN 202011586600 A CN202011586600 A CN 202011586600A CN 112608254 A CN112608254 A CN 112608254A
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- RHGKLRLOHDJJDR-BYPYZUCNSA-N L-citrulline Chemical compound NC(=O)NCCC[C@H]([NH3+])C([O-])=O RHGKLRLOHDJJDR-BYPYZUCNSA-N 0.000 title claims abstract description 179
- 229960002173 citrulline Drugs 0.000 title claims abstract description 93
- 238000000034 method Methods 0.000 title abstract description 31
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 35
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 claims abstract description 26
- 239000013078 crystal Substances 0.000 claims abstract description 25
- 238000001914 filtration Methods 0.000 claims abstract description 19
- 239000012528 membrane Substances 0.000 claims abstract description 19
- 238000006243 chemical reaction Methods 0.000 claims abstract description 17
- 238000001816 cooling Methods 0.000 claims abstract description 14
- 238000001291 vacuum drying Methods 0.000 claims abstract description 8
- 238000004042 decolorization Methods 0.000 claims abstract description 7
- 238000000502 dialysis Methods 0.000 claims abstract description 7
- 238000000967 suction filtration Methods 0.000 claims abstract description 7
- 238000000909 electrodialysis Methods 0.000 claims description 26
- 239000007788 liquid Substances 0.000 claims description 15
- 238000002425 crystallisation Methods 0.000 claims description 13
- 230000008025 crystallization Effects 0.000 claims description 13
- 239000013505 freshwater Substances 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 108010082340 Arginine deiminase Proteins 0.000 claims description 7
- 238000001035 drying Methods 0.000 claims description 7
- 239000004475 Arginine Substances 0.000 claims description 5
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 5
- 238000010612 desalination reaction Methods 0.000 claims description 5
- 238000004140 cleaning Methods 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 abstract description 8
- 230000004151 fermentation Effects 0.000 abstract description 8
- 108090000790 Enzymes Proteins 0.000 abstract description 4
- 102000004190 Enzymes Human genes 0.000 abstract description 4
- 230000000813 microbial effect Effects 0.000 abstract description 4
- 238000003786 synthesis reaction Methods 0.000 abstract description 4
- 230000007613 environmental effect Effects 0.000 abstract description 2
- 239000003344 environmental pollutant Substances 0.000 abstract description 2
- 231100000719 pollutant Toxicity 0.000 abstract description 2
- 238000004886 process control Methods 0.000 abstract description 2
- 238000011033 desalting Methods 0.000 abstract 1
- 239000000047 product Substances 0.000 description 15
- 230000008569 process Effects 0.000 description 9
- RHGKLRLOHDJJDR-UHFFFAOYSA-N Ndelta-carbamoyl-DL-ornithine Natural products OC(=O)C(N)CCCNC(N)=O RHGKLRLOHDJJDR-UHFFFAOYSA-N 0.000 description 7
- 235000013477 citrulline Nutrition 0.000 description 7
- 238000000605 extraction Methods 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 235000009697 arginine Nutrition 0.000 description 4
- 238000004064 recycling Methods 0.000 description 4
- 238000006911 enzymatic reaction Methods 0.000 description 3
- 238000005374 membrane filtration Methods 0.000 description 3
- 238000001471 micro-filtration Methods 0.000 description 3
- 239000012452 mother liquor Substances 0.000 description 3
- 238000001728 nano-filtration Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 2
- 235000014852 L-arginine Nutrition 0.000 description 2
- 229930064664 L-arginine Natural products 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000003287 optical effect Effects 0.000 description 2
- 239000011148 porous material Substances 0.000 description 2
- 238000011084 recovery Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000010865 sewage Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000003440 toxic substance Substances 0.000 description 2
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N valeric acid Chemical compound CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- 241000219109 Citrullus Species 0.000 description 1
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 description 1
- RHGKLRLOHDJJDR-SCSAIBSYSA-N D-citrulline Chemical compound OC(=O)[C@H](N)CCCNC(N)=O RHGKLRLOHDJJDR-SCSAIBSYSA-N 0.000 description 1
- 108010093096 Immobilized Enzymes Proteins 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 150000001370 alpha-amino acid derivatives Chemical class 0.000 description 1
- 235000008206 alpha-amino acids Nutrition 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000003054 catalyst Substances 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000006329 citrullination Effects 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 230000009123 feedback regulation Effects 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000001822 immobilized cell Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000012805 post-processing Methods 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 102220305863 rs1015663503 Human genes 0.000 description 1
- 210000001082 somatic cell Anatomy 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 231100000167 toxic agent Toxicity 0.000 description 1
- 230000004143 urea cycle Effects 0.000 description 1
- 229940005605 valeric acid Drugs 0.000 description 1
- 230000024883 vasodilation Effects 0.000 description 1
- 239000002351 wastewater Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C273/00—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
- C07C273/18—Preparation of urea or its derivatives, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups of substituted ureas
- C07C273/189—Purification, separation, stabilisation, use of additives
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
Abstract
The invention discloses a method for preparing L-citrulline, which comprises the following steps: filtering the conversion solution containing the L-citrulline by an ultrafiltration membrane to obtain ultrafiltration clear solution containing the L-citrulline; step 2: desalting the ultrafiltration clear solution to obtain a dialysis clear solution; and step 3: adjusting pH of the dialyzed clear solution to 4.2-4.6, and adding activated carbon into the dialyzed clear solution for decolorization; and 4, step 4: concentrating the decolorized solution at vacuum degree to obtain a concentrated solution containing L-citrulline 60-70%; and 5: cooling the concentrated solution to 10 ℃, adding seed crystals, cooling and crystallizing for 3-4h, and performing suction filtration to obtain crystals of L-citrulline; step 6: and (3) carrying out vacuum drying on the crystals of the L-citrulline to obtain a purified L-citrulline product. Compared with the traditional chemical synthesis method, the enzyme method and the microbial fermentation method, the method for preparing the L-citrulline has the advantages of simpler process control, no generation of a large amount of pollutants in the production process and environmental friendliness.
Description
Technical Field
The invention relates to the technical field of bioengineering, in particular to a method for preparing L-citrulline.
Background
L-Citrulline (L-Citrulline) with the chemical name of L- (+) -2-amino-5-urea valeric acid and the molecular formula of CAS No 372-75-8C6H13N3O3Molecular weight of 175.19, melting point of 222 ℃, isoelectric point of 5.92, and specific optical rotation [ alpha ]]D25= +24.5 to +26.8 degrees, white crystal or crystalline powder, sour, soluble in water, insoluble in ethanol and ether. L-citrulline is an alpha amino acid named from the watermelon from which citrulline was first extracted.
L-citrulline is an important non-essential amino acid, is an important intermediate in the urea cycle process, and has a plurality of important physiological functions such as free radical removal, foreign body rejection effect indicator, vasodilation, blood pressure stabilization, rheumatoid arthritis diagnosis, antioxidation and the like; in addition, it has very obvious functions of restoring energy and raising immunity of human body. In recent years, L-citrulline is more and more emphasized in the fields of food, cosmetics, medicines and the like, and has wide development and application prospects.
At present, the main production methods of L-citrulline include chemical synthesis methods, enzymatic methods and microbial fermentation methods.
(1) The chemical method means that L-arginine is hydrolyzed under alkaline condition to obtain L-citrulline, the process is difficult to control, the product contains optical antipode D-citrulline, the product quality is influenced, a large amount of waste water is generated in the production process, and the environment is greatly polluted.
(2) The difficulty of the fermentation production is that the yield of the L-citrulline in unit volume is low due to the feedback inhibition in the citrulline metabolic pathway, so the cost for extracting the L-citrulline from the fermentation liquor is high, and the sewage quantity is large. The fermentation method has the advantages of low cost, high yield, high product purity, no toxic substances in the production process of the product, and convenience for post-processing of the product. At present, the country most studied for the production of citrulline by fermentation is japan, and a certain level of fermentation has been achieved.
(3) An enzyme method comprises the following steps: under the action of arginine deiminase, L-arginine is converted into L-citrulline, the production condition is mild, no toxic substance is generated, impurities in a conversion system are less, the extraction process is simple, and the pollution is less. The method takes microbial somatic cells with strong specificity and high conversion rate as a catalyst to catalyze the deimination of arginine to generate L-citrulline. The enzymatic method is used for synthesizing the citrulline, and a one-step enzymatic reaction is mostly adopted, so that the complex feedback regulation effect in the complete synthesis path of the citrulline can be avoided, and the citrulline can be accumulated to a higher concentration. The immobilized enzyme or immobilized cell is adopted for catalysis, so that the difficulty of subsequent separation and purification is further reduced.
However, in the process of separating and purifying the L-citrulline from the enzyme conversion solution, arginine cannot be completely converted, and a large amount of ammonium salt byproducts are generated in the conversion solution, which has a certain influence on the product purity. The existing process has complex extraction process and multiple steps, some of the processes need to use ion exchange resin for product separation, the sewage production amount is large, and the environmental pressure maintaining capacity is high. The citrulline has high solubility in water phase, so the crystallization temperature is low, about 4 ℃, the production has strict requirements on temperature conditions, and the crystallization yield is low.
Disclosure of Invention
The present invention is directed to a method for producing L-citrulline, which solves the problems set forth in the background art.
In order to achieve the purpose, the invention provides the following technical scheme: a method for producing L-citrulline, comprising the following steps:
step 1: and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, recovering and utilizing arginine deiminase to obtain ultrafiltration clear solution containing L-citrulline, and cleaning the ultrafiltration clear solution by using a small amount of pure water;
step 2: electrodialysis: carrying out electrodialysis treatment on the washed ultrafiltration clear liquid of the L-citrulline to further desalt and residue arginine, and obtaining dialysis clear liquid;
and step 3: activated carbon decolorization: adjusting pH of the dialyzed clear solution to 4.2-4.6, adding activated carbon into the dialyzed clear solution, decolorizing at 50-60 deg.C for 25-35min, and filtering with filter membrane to obtain decolorized solution containing L-citrulline;
and 4, step 4: concentration: concentrating decolorized solution at 50-55 deg.C under vacuum degree of 0.095Mpa or more to obtain concentrated solution containing L-citrulline 60-70%;
and 5: and (3) crystallization: cooling the concentrated solution to 10 ℃, adding seed crystals, cooling and crystallizing for 3-4h, and performing suction filtration to obtain crystals of L-citrulline;
step 6: and (3) drying: and (3) carrying out vacuum drying on the crystals of the L-citrulline to obtain a purified L-citrulline product.
Preferably, in the step 1, when the ultrafiltration membrane is used for filtration, the filtration temperature is less than 50 ℃, and the membrane aperture of the ultrafiltration membrane is 3000-10000D.
Preferably, in step 2, the voltage during electrodialysis is 5V, the current during electrodialysis is changed under the condition of constant voltage, the current is changed within the range of 0.1-1A, the temperature is 20-30 ℃, the pH value of the fresh water chamber is maintained at 5.8-6.0 during desalination, and the final fresh water conductivity is 1-5 ms/cm.
Preferably, in step 3, the volume ratio of the dialyzed clear solution to the activated carbon is 1: 0.5-1%.
Preferably, in step 3, a 0.22 μm filter is used for filtration.
Preferably, in step 5, the volume ratio of the concentrated solution to the seed crystal is 1: 0.1-0.5%.
Preferably, in step 6, the temperature for vacuum drying is 50-60 ℃.
Compared with the prior art, the invention has the beneficial effects that:
compared with the traditional chemical synthesis method, the enzyme method and the microbial fermentation method, the method for preparing the L-citrulline has the advantages that the process control is simpler, a large amount of pollutants cannot be generated in the production process, and the method is environment-friendly; meanwhile, ethanol is not used in the production process, and the concentrated crystal is directly crystallized under certain process conditions, so that the method has the advantages of mild production conditions, short concentration time, high product purity, high crystallization yield, high yield, low production cost and the like.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The invention provides a technical scheme that: a method for producing L-citrulline, comprising the following steps:
example 1:
the novel extraction process of the L-citrulline comprises the following steps:
1) and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, namely arginine deiminase (purchased) and recycling to obtain ultrafiltration clear solution containing L-citrulline, and washing a small amount of pure water in the system. And (3) ultrafiltration membrane filtration conditions: the temperature is 10 ℃, and the pore diameter is 3000D;
2) electrodialysis: performing electrodialysis treatment on the microfiltration clear liquid of the L-citrulline for one-step desalination to obtain dialysis clear liquid; electrodialysis conditions: the voltage is 5V, the current of the electrodialysis equipment is a changing process but not constant under the condition of constant voltage, the changing range of the current is 0.1-1A, the temperature is 20 ℃, the pH value of the fresh water chamber is maintained at 5.8, and the final fresh water conductivity is 1 ms/cm;
3) activated carbon decolorization: adjusting pH of the nanofiltration clear liquid to 4.2, adding active carbon according to 0.5% of volume ratio, decolorizing at 50 deg.C for 25 min; filtering with 0.22 μm filter membrane to obtain decolorized solution containing L-citrulline;
4) concentration and crystallization: concentrating the pre-concentrated solution at a vacuum degree of not less than 0.095Mpa and a concentration temperature of 50 ℃ to obtain a concentrated solution with the L-citrulline content of 60-70%; cooling the concentrated solution to 10 ℃, adding 0.1% of seed crystal, cooling and crystallizing for 3h, and performing suction filtration to obtain a crystal of L-citrulline; the crystallization mother liquor returns to electrodialysis circulation for recovery;
5) and (3) drying: and (3) carrying out vacuum drying on the crystals of the L-citrulline at 50 ℃ to obtain a purified L-citrulline product.
Example 2:
the novel extraction process of the L-citrulline comprises the following steps:
1) and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, namely arginine deiminase (purchased) and recycling to obtain ultrafiltration clear solution containing L-citrulline, and washing a small amount of pure water in the system. And (3) ultrafiltration membrane filtration conditions: temperature 20 ℃, aperture 5000D;
2) electrodialysis: performing electrodialysis treatment on the microfiltration clear liquid of the L-citrulline for one-step desalination to obtain dialysis clear liquid; electrodialysis conditions: the voltage is 5V, the current of the electrodialysis equipment is a changing process but not constant under the condition of constant voltage, the changing range of the current is 0.1-1A, the temperature is 20 ℃, the pH value of the fresh water chamber is maintained at 5.8, and the final fresh water conductivity is 1 ms/cm;
3) activated carbon decolorization: adjusting pH of the nanofiltration clear liquid to 4.5, adding active carbon according to 0.7% of volume ratio, decolorizing at 55 deg.C for 30 min; filtering with 0.22 μm filter membrane to obtain decolorized solution containing L-citrulline;
4) concentration and crystallization: concentrating the pre-concentrated solution at a vacuum degree of not less than 0.096Mpa and a concentration temperature of 50 ℃ to obtain a concentrated solution with the L-citrulline content of 60-70%; cooling the concentrated solution to 10 ℃, adding 0.3% of seed crystal, cooling and crystallizing for 3.5h, and performing suction filtration to obtain a crystal of L-citrulline; the crystallization mother liquor returns to electrodialysis circulation for recovery;
5) and (3) drying: and drying the crystals of the L-citrulline at 55 ℃ in vacuum to obtain a purified L-citrulline product.
Example 3:
the novel extraction process of the L-citrulline comprises the following steps:
1) and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, namely arginine deiminase (purchased) and recycling to obtain ultrafiltration clear solution containing L-citrulline, and washing a small amount of pure water in the system. And (3) ultrafiltration membrane filtration conditions: the temperature is 40 ℃, and the pore diameter is 10000D;
2) electrodialysis: performing electrodialysis treatment on the microfiltration clear liquid of the L-citrulline for one-step desalination to obtain dialysis clear liquid; electrodialysis conditions: the voltage is 5V, the current of the electrodialysis equipment is a changing process but not constant under the condition of constant voltage, the changing range of the current is 0.1-1A, the temperature is 20 ℃, the pH value of the fresh water chamber is maintained at 5.8, and the final fresh water conductivity is 1 ms/cm;
3) activated carbon decolorization: adjusting the pH value of the nanofiltration clear liquid to 4.6, adding activated carbon according to 1% of the volume ratio, and decoloring at 60 ℃ for about 35 min; filtering with 0.22 μm filter membrane to obtain decolorized solution containing L-citrulline;
4) concentration and crystallization: concentrating the pre-concentrated solution at a vacuum degree of more than or equal to 0.098Mpa and a concentration temperature of 55 ℃ to obtain a concentrated solution with the L-citrulline content of 60-70%; cooling the concentrated solution to 10 ℃, adding 0.5% of seed crystal, cooling and crystallizing for 4h, performing suction filtration to obtain crystals of L-citrulline, and returning the crystallization mother liquor to electrodialysis for recycling;
5) and (3) drying: and (3) carrying out vacuum drying on the crystals of the L-citrulline at 60 ℃ to obtain a purified L-citrulline product.
In the three embodiments, the yield of ultrafiltration and electrodialysis reaches 99%, the comprehensive yield of the concentrated and crystallized product is more than 90%, and the method has the advantages of mild production conditions, short concentration time, high product purity, high crystallization yield, high yield, low production cost and the like.
Application method
A method for producing L-citrulline, comprising the following steps:
and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, recovering and utilizing arginine deiminase to obtain ultrafiltration clear solution containing L-citrulline, and cleaning the ultrafiltration clear solution by using a small amount of pure water;
step 2: electrodialysis: carrying out electrodialysis treatment on the washed ultrafiltration clear liquid of the L-citrulline to further desalt and residue arginine, and obtaining dialysis clear liquid;
and step 3: activated carbon decolorization: adjusting pH of the dialyzed clear solution to 4.2-4.6, adding activated carbon into the dialyzed clear solution, decolorizing at 50-60 deg.C for 25-35min, and filtering with filter membrane to obtain decolorized solution containing L-citrulline;
and 4, step 4: concentration: concentrating decolorized solution at 50-55 deg.C under vacuum degree of 0.095Mpa or more to obtain concentrated solution containing L-citrulline 60-70%;
and 5: and (3) crystallization: cooling the concentrated solution to 10 ℃, adding seed crystals, cooling and crystallizing for 3-4h, and performing suction filtration to obtain crystals of L-citrulline;
step 6: and (3) drying: and (3) carrying out vacuum drying on the crystals of the L-citrulline to obtain a purified L-citrulline product.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (7)
1. A method for producing L-citrulline, which is characterized by comprising the following steps:
step 1: and (3) ultrafiltration: filtering the conversion solution containing L-citrulline by an ultrafiltration membrane, removing the conversion solution, recovering and utilizing arginine deiminase to obtain ultrafiltration clear solution containing L-citrulline, and cleaning the ultrafiltration clear solution by using a small amount of pure water;
step 2: electrodialysis: carrying out electrodialysis treatment on the washed ultrafiltration clear liquid of the L-citrulline to further desalt and residue arginine, and obtaining dialysis clear liquid;
and step 3: activated carbon decolorization: adjusting pH of the dialyzed clear solution to 4.2-4.6, adding activated carbon into the dialyzed clear solution, decolorizing at 50-60 deg.C for 25-35min, and filtering with filter membrane to obtain decolorized solution containing L-citrulline;
and 4, step 4: concentration: concentrating decolorized solution at 50-55 deg.C under vacuum degree of 0.095Mpa or more to obtain concentrated solution containing L-citrulline 60-70%;
and 5: and (3) crystallization: cooling the concentrated solution to 10 ℃, adding seed crystals, cooling and crystallizing for 3-4h, and performing suction filtration to obtain crystals of L-citrulline;
step 6: and (3) drying: and (3) carrying out vacuum drying on the crystals of the L-citrulline to obtain a purified L-citrulline product.
2. The method for producing L-citrulline according to claim 1, characterized in that: in the step 1, when the ultrafiltration membrane is used for filtration, the filtration temperature is less than 50 ℃, and the filter membrane aperture of the ultrafiltration membrane is 3000-10000D.
3. The method for producing L-citrulline according to claim 1, characterized in that: in step 2, the voltage during electrodialysis is 5V, the current of electrodialysis is changed under the condition of constant voltage, the change range of the current is 0.1-1A, the temperature is 20-30 ℃, the pH value of a fresh water chamber is maintained at 5.8-6.0 during desalination, and the final fresh water conductivity is 1-5 ms/cm.
4. The method for producing L-citrulline according to claim 1, characterized in that: in step 3, the volume ratio of the dialyzed clear solution to the activated carbon is 1: 0.5-1%.
5. The method for producing L-citrulline according to claim 1, characterized in that: in step 3, a 0.22 μm filter was used for filtration.
6. The method for producing L-citrulline according to claim 1, characterized in that: in step 5, the volume ratio of the concentrated solution to the seed crystal is 1: 0.1-0.5%.
7. The method for producing L-citrulline according to claim 1, characterized in that: in step 6, the temperature of vacuum drying is 50-60 ℃.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN113881724A (en) * | 2021-09-30 | 2022-01-04 | 新泰市佳禾生物科技有限公司 | Extraction and purification method for arginine-citrulline |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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