CN1789236A - Purification method of 15N-L-arginine - Google Patents
Purification method of 15N-L-arginine Download PDFInfo
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- CN1789236A CN1789236A CN 200410089567 CN200410089567A CN1789236A CN 1789236 A CN1789236 A CN 1789236A CN 200410089567 CN200410089567 CN 200410089567 CN 200410089567 A CN200410089567 A CN 200410089567A CN 1789236 A CN1789236 A CN 1789236A
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Abstract
The invention discloses a method for separating and purifying15N-L-arginine, employing the fermentation liquor containing arginine as raw material, removing the coloring matter and foreign matter in the fermentation liquor with strong acidic resin, eluting with ammonial solution, separating the arginine from other amino acid, decolourizing the vacuum compression liquor with activated char, vacuum compressing the filtering liquid, adding compression for refinishing, and getting the 15N-L-arginine with the purity >98%. The separating and purifying method is characterized by simple process, convenient operation and more than 80% of the total extraction rate.
Description
Technical field
The present invention relates to a kind of amino acid whose separating and purifying method, particularly a kind of
15The arginic separating and purifying method of N-L-.
Background technology
The L-arginine is the intravital semi-dispensable amino acid of human body and animal, also is a kind of important mesostate of organism ornithine cycle, in fields such as medicine industry, foodstuffs industry important purposes is arranged.
15The N-L-arginine is widely used as tracer agent in research fields such as medical science, life sciences owing to the singularity of its structure.
15Biological fermentation process or proteolysis method are generally adopted in the arginic preparation of N-L-.No matter be which kind of method is produced arginine, all to relate to the problem that arginine and a large amount of impurity are separated.
Prior art is separated arginine and is generally adopted the precipitator method, electroosmose process and ion exchange method.
The precipitator method are to utilize precipitation agent (as phenyl aldehyde, Pentachlorophenol, dodecyl semi-annular jade pendant acid sodium etc.) can make the less mixture of arginine and its generation solubleness and precipitate, thereby reach arginine and the isolating purpose of other amino acid.But the precipitation agent phenyl aldehyde in this method, Pentachlorophenol are toxic compounds, and the discharging of waste liquid behind the extraction arginine can pollute environment; Though and the toxicological harmless of dodecyl semi-annular jade pendant acid sodium, this method complex operation step, dosage is wayward.
15The tracer agent that the N-L-arginine is used as industries such as medicine, food should not adopt above three kinds of solvents to carry out purifying.
Though the electroosmose process principle is simple, this method investment is bigger, and the arginine rate of recovery is not high, thereby it is wideless to make this method use.
It is more that ion exchange method is extracted arginic research report, but do not form large-scale production so far yet.At present, there is a large amount of impurity (mainly being pigment and multiple other amino acid) in this method owing to containing in the arginic mixed solution, thereby has influenced exchange capacity, can not separate with other amino acid during wash-out, has influenced total extract yield.
Summary of the invention
Purpose of the present invention is to overcome the deficiency that prior art exists, and provides a kind of new
15The arginic separating and purifying method of N-L-, this method directly separate from bio-fermented liquid purifies
15The N-L-arginine, thus poisonous operation avoided, improved
15The arginic yield of N-L-.
To achieve these goals, the present invention has adopted following technical scheme; A kind of
15The arginic separating and purifying method of N-L-is a raw material to contain arginic fermented liquid, takes following steps to separate and purifies:
A, will contain arginic fermented liquid and be adjusted to acidity with oxalic acid, centrifugation goes out supernatant liquor, and supernatant liquor adsorbs with highly acidic resin, uses the ammoniacal liquor desorb, is contained
15The arginic stripping liquid of N-L-;
B, step a gained is contained
15The arginic stripping liquid vacuum concentration of N-L-behind dissolved in distilled water, adds activated carbon decolorizing to doing, and suction filtration is contained
15The arginic clear filtrate of N-L-;
C, step b gained is contained
15The arginic clear filtrate vacuum concentration of N-L-adds dehydrated alcohol, obtains purified after crystallisation by cooling, filtration, vacuum-drying
15N-L-arginine product.
Described in the step a to be adjusted to tart pH value with oxalic acid be 3-4, described highly acidic resin is that gel strong acid benzene hexene is a resin, described desorb ammonia concn is 0.05～0.2mol/L.
Decolouring described in the step b is carried out under 70～90 ℃.
Temperature during adding dehydrated alcohol described in the step c is controlled at 60～80 ℃, and the temperature during described crystallisation by cooling is controlled at room temperature to-18 ℃, and the temperature during described vacuum-drying is controlled at 50～60 ℃.
Void column flow velocity SV=0.1～1.2% of the resin absorption control centrifuged supernatant described in the step a.
The present invention
15The arginic separating and purifying method of N-L-makes it compared with prior art owing to adopted above technical scheme, has the following advantages and characteristics:
(1) avoided nontoxic operation, fermented liquid need not to carry out pre-treatment, and adopting gel strong acid benzene hexene is that resin can be removed a large amount of pigments in the fermented liquid, and can make
15The N-L-arginine separates with other amino acid (as Threonine, proline(Pro) etc.),
15The arginic single spot yield of N-L-＞90%, therefore, this technology was both simple, was convenient to operation again.
(2) use dehydrated alcohol as refining solvent, pigment can be stayed in the mother liquor morely, the product purity that makes＞98%, color and luster is snow-white,
15The arginic total extract yield of N-L-＞80%.
Embodiment
Below by several specific embodiments to the present invention
15The arginic separating and purifying method of N-L-is further described.
Embodiment 1
What obtain after will fermenting with Corynebacterium glutamicum contains
15The arginic fermented liquid of N-L-is regulated pH=3.0 with oxalic acid, remove mycelium and throw out in the fermented liquid with the whizzer centrifugation, obtain supernatant liquor, is resin column absorption with supernatant liquor by gel strong acid benzene hexene, control void column flow velocity SV=0.3%, clean resin to neutral with distilled water afterwards, use the ammoniacal liquor desorb of 0.10mol/L then, only contained
15The arginic stripping liquid of N-L-.Stripping liquid adds an amount of dissolved in distilled water after vacuum concentration is extremely done, add activated carbon decolorizing down at 75 ℃, and the clear filtrate behind the suction filtration is again through vacuum concentration, and the adding dehydrated alcohol is cooled to room temperature then under 70 ℃, in-18 ℃ of freeze overnight,
15The crystallization of N-L-arginine is separated out, and crystallization is filtered, drained together with mother liquor, with absolute ethanol washing twice, refilters, drains, and obtains
15The arginic crystallisate of N-L-, then with crystallisate in 60 ℃ of vacuum-dryings, obtain purified
15N-L-arginine product.
Embodiment 2
What obtain after will fermenting with Corynebacterium glutamicum contains
15The arginic fermented liquid of N-L-is regulated pH=3.6 with oxalic acid, remove mycelium and throw out in the fermented liquid with the whizzer centrifugation, obtain supernatant liquor, is resin column absorption with supernatant liquor by gel strong acid benzene hexene, control void column flow velocity SV=0.8%, clean resin to neutral with distilled water afterwards, use the ammoniacal liquor desorb of 0.15mol/L then, only contained
15The arginic stripping liquid of N-L-.The stripping liquid vacuum concentration adds an amount of dissolved in distilled water after extremely doing, and adds activated carbon decolorizing down at 80 ℃, and the clear filtrate behind the suction filtration in 80 ℃ of adding dehydrated alcohols, is cooled to room temperature again through vacuum concentration, in-18 ℃ of freeze overnight,
15The crystallization of N-L-arginine is separated out, and crystallization is filtered, drained together with mother liquor, with absolute ethanol washing twice, refilters, drains, and obtains
15The arginic crystallisate of N-L-, then with crystallisate in 50 ℃ of vacuum-dryings, obtain purified
15N-L-arginine product.
Embodiment 3
What obtain after will fermenting with Corynebacterium glutamicum contains
15The arginic fermented liquid of N-L-is regulated pH=4 with oxalic acid, remove mycelium and throw out in the fermented liquid with the whizzer centrifugation, obtain supernatant liquor, is resin column absorption with supernatant liquor by gel strong acid benzene hexene, control void column flow velocity SV=1.2%, clean resin to neutral with distilled water afterwards, use the ammoniacal liquor desorb of 0.2mol/L then, only contained
15The arginic stripping liquid of N-L-.The stripping liquid vacuum concentration adds an amount of dissolved in distilled water after extremely doing, and adds activated carbon decolorizing down at 90 ℃, and the clear filtrate behind the suction filtration in 60 ℃ of adding dehydrated alcohols, is cooled to room temperature again through vacuum concentration, in-18 ℃ of freeze overnight,
15The crystallization of N-L-arginine is separated out, and crystallization is filtered, drained together with mother liquor, with absolute ethanol washing twice, refilters, drains, and obtains
15The arginic crystallisate of N-L-, then with crystallisate in 55 ℃ of vacuum-dryings, obtain purified
15N-L-arginine product.
In the foregoing description, gained
15The purity of N-L-arginine product reaches more than 98%, always extracts yield and reaches more than 80%.When adopting N atom abundance is 5.31% low abundance feedstock production
15During the N-L-arginine, in the products obtained therefrom
15The N abundance reaches more than 5.1%, is 98.90% high abundance feedstock production when employing N atom abundance
15During the N-L-arginine, in the products obtained therefrom
15The N abundance reaches more than 98%.
Claims (5)
1. one kind
15The arginic separating and purifying method of N-L-is characterized in that: to contain arginic fermented liquid is raw material, takes following steps to separate and purifies:
A, will contain arginic fermented liquid and be adjusted to acidity with oxalic acid, centrifugation goes out supernatant liquor, and supernatant liquor adsorbs with highly acidic resin, uses the ammoniacal liquor desorb, is contained
15The arginic stripping liquid of N-L-;
B, step a gained is contained
15The arginic stripping liquid vacuum concentration of N-L-behind dissolved in distilled water, adds activated carbon decolorizing to doing, and suction filtration is contained
15The arginic clear filtrate of N-L-;
C, step b gained is contained
15The arginic clear filtrate vacuum concentration of N-L-adds dehydrated alcohol, obtains purified after crystallisation by cooling, filtration, vacuum-drying
15N-L-arginine product.
2. as claimed in claim 1
15The arginic separating and purifying method of N-L-is characterized in that: described in the step a to be adjusted to tart pH value with oxalic acid be 3-4, described highly acidic resin is that gel strong acid benzene hexene is a resin, described desorb ammonia concn is 0.05～0.2mol/L.
3. as claimed in claim 1
15The arginic separating and purifying method of N-L-is characterized in that: the decolouring described in the step b is carried out under 70～90 ℃.
4. as claimed in claim 1
15The arginic separating and purifying method of N-L-, it is characterized in that: the temperature during adding dehydrated alcohol described in the step c is controlled at 60～80 ℃, temperature during described crystallisation by cooling is controlled at room temperature to-18 ℃, and the temperature during described vacuum-drying is controlled at 50～60 ℃.
5. as claimed in claim 1
15The arginic separating and purifying method of N-L-is characterized in that: void column flow velocity SV=0.1～1.2% of the resin absorption control centrifuged supernatant described in the step a.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100895677A CN100509757C (en) | 2004-12-15 | 2004-12-15 | Purification method of *N-L-arginine |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2004100895677A CN100509757C (en) | 2004-12-15 | 2004-12-15 | Purification method of *N-L-arginine |
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| Publication Number | Publication Date |
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| CN1789236A true CN1789236A (en) | 2006-06-21 |
| CN100509757C CN100509757C (en) | 2009-07-08 |
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| Application Number | Title | Priority Date | Filing Date |
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| CNB2004100895677A Expired - Fee Related CN100509757C (en) | 2004-12-15 | 2004-12-15 | Purification method of *N-L-arginine |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102001972A (en) * | 2010-10-26 | 2011-04-06 | 广东肇庆星湖生物科技股份有限公司 | Method for separating and extracting L-arginine from fermentation liquor |
| CN102972636A (en) * | 2012-12-03 | 2013-03-20 | 江苏赛奥生化有限公司 | Manufacturing method of feed-grade L-arginine premixing agent |
| CN103667381A (en) * | 2013-12-24 | 2014-03-26 | 山东民强生物科技股份有限公司 | Method for improving yield of arginine |
| CN103695489A (en) * | 2013-12-24 | 2014-04-02 | 山东民强生物科技股份有限公司 | Arginine refining technology |
| CN105732436A (en) * | 2016-04-20 | 2016-07-06 | 上海化工研究院 | Method for extracting high-abundance L-arginine-15N4 from high-abundance 15N isotope-labeled L-arginine fermentation liquor |
| CN112479935A (en) * | 2020-12-03 | 2021-03-12 | 江苏优普生物化学科技股份有限公司 | Arginine purification and refining process |
-
2004
- 2004-12-15 CN CNB2004100895677A patent/CN100509757C/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102001972A (en) * | 2010-10-26 | 2011-04-06 | 广东肇庆星湖生物科技股份有限公司 | Method for separating and extracting L-arginine from fermentation liquor |
| CN102972636A (en) * | 2012-12-03 | 2013-03-20 | 江苏赛奥生化有限公司 | Manufacturing method of feed-grade L-arginine premixing agent |
| CN103667381A (en) * | 2013-12-24 | 2014-03-26 | 山东民强生物科技股份有限公司 | Method for improving yield of arginine |
| CN103695489A (en) * | 2013-12-24 | 2014-04-02 | 山东民强生物科技股份有限公司 | Arginine refining technology |
| CN103695489B (en) * | 2013-12-24 | 2015-09-02 | 山东民强生物科技股份有限公司 | A kind of arginine process for refining |
| CN103667381B (en) * | 2013-12-24 | 2015-09-30 | 山东民强生物科技股份有限公司 | A kind of method improving yield of arginine |
| CN105732436A (en) * | 2016-04-20 | 2016-07-06 | 上海化工研究院 | Method for extracting high-abundance L-arginine-15N4 from high-abundance 15N isotope-labeled L-arginine fermentation liquor |
| CN112479935A (en) * | 2020-12-03 | 2021-03-12 | 江苏优普生物化学科技股份有限公司 | Arginine purification and refining process |
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| Publication number | Publication date |
|---|---|
| CN100509757C (en) | 2009-07-08 |
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Owner name: SHANGHAI LIANQI CHEMICAL TECHNOLOGY CO., LTD. |
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Effective date of registration: 20100617 Address after: 200062 Shanghai Yunling Road No. 345 Co-patentee after: SHANGHAI LIANQI CHEMICAL SCIENCE & TECHNOLOGY CO., LTD. Patentee after: Shanghai Research Institute of Chemical Industry Address before: 200062 Shanghai Yunling Road No. 345 Patentee before: Shanghai Research Institute of Chemical Industry |
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Granted publication date: 20090708 Termination date: 20101215 |