CN106278954A - A kind of method extracting agmatine - Google Patents
A kind of method extracting agmatine Download PDFInfo
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- CN106278954A CN106278954A CN201610684104.8A CN201610684104A CN106278954A CN 106278954 A CN106278954 A CN 106278954A CN 201610684104 A CN201610684104 A CN 201610684104A CN 106278954 A CN106278954 A CN 106278954A
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- Prior art keywords
- agmatine
- clear liquid
- method extracting
- arginine
- thalline
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- QYPPJABKJHAVHS-UHFFFAOYSA-N Agmatine Natural products NCCCCNC(N)=N QYPPJABKJHAVHS-UHFFFAOYSA-N 0.000 title claims abstract description 38
- QYPPJABKJHAVHS-UHFFFAOYSA-P agmatinium(2+) Chemical compound NC(=[NH2+])NCCCC[NH3+] QYPPJABKJHAVHS-UHFFFAOYSA-P 0.000 title claims abstract description 38
- 238000000034 method Methods 0.000 title claims abstract description 24
- 239000007788 liquid Substances 0.000 claims abstract description 63
- 238000000502 dialysis Methods 0.000 claims abstract description 23
- 238000006243 chemical reaction Methods 0.000 claims abstract description 22
- 102100032252 Antizyme inhibitor 2 Human genes 0.000 claims abstract description 21
- 101000798222 Homo sapiens Antizyme inhibitor 2 Proteins 0.000 claims abstract description 20
- 238000010257 thawing Methods 0.000 claims abstract description 15
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims abstract description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 11
- 230000000694 effects Effects 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract description 9
- 239000004475 Arginine Substances 0.000 claims abstract description 8
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 8
- 230000002779 inactivation Effects 0.000 claims abstract description 8
- 239000002994 raw material Substances 0.000 claims abstract description 8
- 239000013078 crystal Substances 0.000 claims abstract description 6
- 238000000703 high-speed centrifugation Methods 0.000 claims abstract description 6
- 230000004071 biological effect Effects 0.000 claims abstract description 3
- 229920002521 macromolecule Polymers 0.000 claims abstract description 3
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 3
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 3
- 239000012528 membrane Substances 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 6
- 238000000108 ultra-filtration Methods 0.000 claims description 6
- 239000002738 chelating agent Substances 0.000 claims description 5
- 230000000844 anti-bacterial effect Effects 0.000 claims description 4
- 102000004190 Enzymes Human genes 0.000 claims description 2
- 108090000790 Enzymes Proteins 0.000 claims description 2
- 238000006114 decarboxylation reaction Methods 0.000 claims description 2
- 235000014666 liquid concentrate Nutrition 0.000 claims description 2
- 239000000047 product Substances 0.000 abstract description 10
- 239000012043 crude product Substances 0.000 abstract description 8
- 239000012535 impurity Substances 0.000 abstract description 6
- 230000008901 benefit Effects 0.000 abstract description 5
- 230000007423 decrease Effects 0.000 abstract description 5
- 239000007806 chemical reaction intermediate Substances 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 238000012545 processing Methods 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 2
- 239000012141 concentrate Substances 0.000 abstract 1
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 10
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000004061 bleaching Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 230000036541 health Effects 0.000 description 3
- 230000008676 import Effects 0.000 description 3
- 238000009413 insulation Methods 0.000 description 3
- 238000007086 side reaction Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 230000001629 suppression Effects 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 2
- HQABUPZFAYXKJW-UHFFFAOYSA-N butan-1-amine Chemical compound CCCCN HQABUPZFAYXKJW-UHFFFAOYSA-N 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000543 intermediate Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- BQJCRHHNABKAKU-KBQPJGBKSA-N morphine Chemical compound O([C@H]1[C@H](C=C[C@H]23)O)C4=C5[C@@]12CCN(C)[C@@H]3CC5=CC=C4O BQJCRHHNABKAKU-KBQPJGBKSA-N 0.000 description 2
- 239000008213 purified water Substances 0.000 description 2
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 2
- 238000002834 transmittance Methods 0.000 description 2
- 238000010792 warming Methods 0.000 description 2
- 239000002912 waste gas Substances 0.000 description 2
- PTAYFGHRDOMJGC-UHFFFAOYSA-N 4-aminobutyl(diaminomethylidene)azanium;hydrogen sulfate Chemical compound OS(O)(=O)=O.NCCCCN=C(N)N PTAYFGHRDOMJGC-UHFFFAOYSA-N 0.000 description 1
- 102000004031 Carboxy-Lyases Human genes 0.000 description 1
- 108090000489 Carboxy-Lyases Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000018997 Growth Hormone Human genes 0.000 description 1
- 108010051696 Growth Hormone Proteins 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 208000007271 Substance Withdrawal Syndrome Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Natural products NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 206010048010 Withdrawal syndrome Diseases 0.000 description 1
- SSBRSHIQIANGKS-UHFFFAOYSA-N [amino(hydroxy)methylidene]azanium;hydrogen sulfate Chemical compound NC(N)=O.OS(O)(=O)=O SSBRSHIQIANGKS-UHFFFAOYSA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 230000004872 arterial blood pressure Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000747 cardiac effect Effects 0.000 description 1
- 210000000748 cardiovascular system Anatomy 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 238000007596 consolidation process Methods 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000000601 hypothalamic hormone Substances 0.000 description 1
- 229940043650 hypothalamic hormone Drugs 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229960005181 morphine Drugs 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 229940124636 opioid drug Drugs 0.000 description 1
- 230000036513 peripheral conductance Effects 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- FYRHIOVKTDQVFC-UHFFFAOYSA-M potassium phthalimide Chemical compound [K+].C1=CC=C2C(=O)[N-]C(=O)C2=C1 FYRHIOVKTDQVFC-UHFFFAOYSA-M 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035943 smell Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000001117 sulphuric acid Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C277/00—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
- C07C277/08—Preparation of guanidine or its derivatives, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups of substituted guanidines
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
Landscapes
- Organic Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention discloses a kind of method extracting agmatine, relate to the extractive technique field of compound.Comprise the following steps: (1) uses raw material to be arginine, and the bioenzymatic conversion feed liquid containing agmatine produced under arginine decarboxylase effect carries out freeze thawing treatment, makes arginine decarboxylase lose biological activity；(2) take the bioenzymatic conversion feed liquid after inactivation and carry out high speed centrifugation, isolate thalline and protein matter, obtain centrifugal clear liquid；(3) centrifugal clear liquid is crossed membranous system, remove thalline relic and macromolecular substances, obtained film dialysis clear liquid；(4) concentrate crossing film dialysis clear liquid, regulate pH after activated carbon decolorizing and carry out solvent crystal, filter to obtain agmatine crude product.This method can quickly process acquisition agmatine crude product, decreases chemical reaction intermediate processing steps, decreases impurity in product, simultaneously, capacity efficiency is high, and low cost is easy and simple to handle, good product quality, improves product quality and yield, creates good economic benefit.
Description
Technical field
The present invention relates to the extractive technique field of compound.
Background technology
Agmatine (gamatine) is the material of a multi-biological function, discharges hypothalamic hormone by stimulating
Carry out work, it is possible to promoting Alfasone and growth hormone, agmatine has multiple benefits, it is possible to increase health durability degree, to motion
Member and bodybuilders have certain benefit and it can be made satisfied, can accelerate physical recovery simultaneously, improve muscle quantities, reduce obesity
Oils and fats, improves health performance, and to pursuing, health is dynamic and to seek each age level healthy lifestyles helpful, guanidine
Base butylamine can also improve nitric oxide production content by two kinds of approach, stimulates release nitric oxide and suppression Nitric oxide syntheses
Synthesis (Nitric oxide syntheses can consume nitric oxide).
Having data to show, central nervous system, gamatine can strengthen the analgesic effect of opioid drug, slow down drug resistance
The generation of property and improve Acute Morphine withdrawal syndrome, gamatine can also promote consolidation of memory in addition.At cardiovascular system,
Gamatine can reduce heart rate, and mean arterial pressure, left chamber pressure, peripheral vascular resistance and cardiac index etc., gamatine is also simultaneously
Gastric acid and pepsic secretion can be increased, make mucosal thickness thinning, deteriorate the mucosa infection etc. that stress cause.There is research simultaneously
Showing, agmatine also has the effect of clear and definite suppression tumor cell proliferation, and the effect of its suppression tumor cell proliferation has tumor
Cell strain specificity.
Present stage, main by chemical synthesis acquisition gamatine on market, primary raw material has Putriscine, O-first
Base isourea sulfate, sulphuric acid, alcohol etc., course of reaction energy consumption is high, and yield is low, and impurity is many, and potential safety hazard is big, has certain dirt to environment
Dye；Also have employing Isosorbide-5-Nitrae-dibromobutane and potassium phthalimide is raw material simultaneously, is substituted, amine solution and guanidinated three steps
Reaction prepares the report of agmatine sulfate, this route reaction mild condition, but side reaction is more, and complex operation, and abnormal smells from the patient is difficult
Hearing, course of reaction uses relatively multi-solvent, does not meets the Production requirement of clean and effective.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of method extracting agmatine, and the method can quickly process and obtain
Obtain agmatine crude product, relatively conventional chemical synthesis, effectively reduce chemical reaction intermediate processing steps, decrease intermediate
Side reaction and during impurity, useless solvent, the generation of waste gas, meanwhile, capacity efficiency is high, and low cost is easy and simple to handle, product matter
Measure, decrease impurity in product, improve product quality and yield, create good economic benefit.
For solving above-mentioned technical problem, the technical solution used in the present invention is: a kind of method extracting agmatine, including
Following steps:
(1) using raw material is arginine, in the lower generation of arginine decarboxylase (arginine decarboxylase, ADC) effect
The bioenzymatic conversion feed liquid containing agmatine carry out freeze thawing treatment, make arginine decarboxylase lose biological activity；
(2) take the bioenzymatic conversion feed liquid after inactivation and carry out high speed centrifugation, isolate thalline and protein matter, obtain centrifugal clear
Liquid；
(3) centrifugal clear liquid is crossed membranous system, remove thalline relic and macromolecular substances, obtained film dialysis clear liquid；
(4) by cross film dialysis clear liquid concentrate, after activated carbon decolorizing regulate pH carry out solvent crystal, filter agmatine is thick
Product.
Preferably, in step (1), arginine decarboxylase is the arginine decarboxylase from antibacterial or plant-derived smart ammonia
Acid decarboxylase.
Preferably, in step (1), freeze thawing treatment, use-20 DEG C～5 DEG C ,-30 DEG C～10 DEG C ,-35 DEG C～20 DEG C successively
Freeze thawing treatment scheme.
It is further preferred that in step (1), freeze thawing treatment, use the mode of filtration to isolate portion after dissolving for the first time
Divide thalline.
Preferably, in step (2), take after the bioenzymatic conversion feed liquid after inactivation adds chelating agent or flocculant, carry out from
The heart.
It is further preferred that chelating agent used or flocculant have: mercaptoethylmaine, polyacrylamide, TGA, thiourea or
EDTA(ethylenediaminetetraacetic acid).
Preferably, in step (2), high speed centrifugation uses disk centrifugal separator or tube centrifuge, centrifugal rotational speed 12000～
15000 revs/min.
Preferably, in step (2), high speed centrifugation uses stacked centrifuge, and centrifugal rotational speed is at 15000 revs/min.
Preferably, in step (3), membranous system is pottery filter membrane and ultrafilter membrane.
It is further preferred that in step (3), pottery filter sizes is 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, one-level fenestra
Footpath 8000-10000 dalton, secondary membrane aperture 3000-5000 dalton.
In step (3), ultrafiltration, secondary membrane aperture can also be 1500-3000 dalton；
Obtain film dialysis clear liquid after two-stage ultrafiltering, cross film dialysis clear liquid light transmittance more than 97%.
Preferably, in step (4), pH is 3.0-7.0.
Preferably, in step (4), after film dialysis clear liquid was concentrated into the 20-50% of film dialysis clear liquid original volume excessively, add
Enter the medicinal carbon of agmatine quality 0.5%-3% in film dialysis clear liquid and carry out adsorption bleaching.
Preferably, in step (4), solvent is methanol, ethanol or isopropanol, and solvent consumption is that the film of crossing after regulation pH is dialysed
1.2-4.5 times of supernatant volume.
Use and have the beneficial effects that produced by technique scheme: the inventive method can quickly process acquisition agmatine
Crude product, relatively conventional chemical synthesis, effectively reduce chemical reaction intermediate processing steps, decrease intermediate side reaction and
During impurity, useless solvent, the generation of waste gas, meanwhile, capacity efficiency is high, and low cost is easy and simple to handle, good product quality, subtracts
Lack impurity in product, improve product quality and yield, create good economic benefit.
Detailed description of the invention
Below in conjunction with detailed description of the invention, the present invention is further detailed explanation.
Embodiment 1
(1) using raw material is arginine, arginine decarboxylase (arginine decarboxylase is the arginine decarboxylase from antibacterial)
Lower bioenzymatic conversion feed liquid 500L containing agmatine 105kg produced of effect is placed in freeze dryer distributor pipe, carries out at freeze thawing
Reason, fast cooling is incubated 1 hour to-20 DEG C, is rapidly heated to 5 DEG C, is directly placed in centrifuge by the liquid in distributor pipe, gets rid of
Filter cutting out partial thalline；Liquid rejection filter gone out the most averagely imports in freeze dryer distributor pipe, fast cooling to-30 DEG C of insulations
It is warming up to 10 DEG C after 1 hour again, is cooled to-35 DEG C in the same way and rises again again to 20 DEG C.
(2) the bioenzymatic conversion feed liquid (freeze thawing inactivation) that will rise again, adds the chelating agent of bioenzymatic conversion feed liquid quality 2%,
Slowly putting into disk centrifugal separator after stirring 1 hour, centrifugal rotational speed, at 12000～15000 revs/min, is isolated thalline, is adopted continuously
Go out clear liquid, obtain centrifugal clear liquid 470L.
(3) above-mentioned clear liquid 470L carries out pottery filter, and pottery filter sizes is 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, one-level
Membrane aperture 8000-10000 dalton, secondary membrane aperture 3000-5000 dalton, two-stage ultrafiltering, after terminating, use 50L purified water
Washing membranous system, obtained film dialysis clear liquid 500L, detected agmatine content 193g/L, yield 92.3%, crossed film dialysis clear liquid printing opacity
Rate is more than 97%.
(4) take above-mentioned cross film dialysis clear liquid 500L be concentrated into film dialysis clear liquid original volume 50% after, add 3kg activity
Charcoal adsorption bleaching, after filtration, regulates pH to 5.0, adds 500L isopropanol and separates out crystal, filter to obtain agmatine in gained feed liquid
Crude product, 65 DEG C of vacuum is dried, detects moisture 0.26%, pure 90.22kg, yield 93.5%.
Embodiment 2
(1) using raw material is arginine, and at arginine decarboxylase, (arginine decarboxylase is from plant-derived arginine decarboxylation
Enzyme) act on and descend the bioenzymatic conversion feed liquid containing agmatine produced to carry out freeze thawing treatment, make arginine decarboxylase lose biology
Activity.Freeze dryer distributor pipe 80 is cooled to-20 DEG C, in every distributor pipe, adds 5L bioenzymatic conversion feed liquid, constant temperature-20
DEG C insulation 1 hour, be rapidly heated to 5 DEG C, the liquid in distributor pipe be directly placed in centrifuge, rejection filter cutting out partial thalline
Amount to 4.6kg；Liquid rejection filter gone out the most averagely imports in distributor pipe, and fast cooling heats up after being incubated 1 hour to-30 DEG C again
To 10 DEG C, be cooled to-35 DEG C in the same way and rise again again to 20 DEG C.
(2) the bioenzymatic conversion feed liquid (freeze thawing inactivation) that will rise again, the bioenzymatic conversion feed liquid quality 3% that addition is risen again
Chelating agent, slowly puts into disk centrifugal separator after stirring 1 hour, and centrifugal rotational speed, at 12000～15000 revs/min, isolates bacterium
Body, continuous extraction clear liquid, obtain centrifugal clear liquid 355L.
(3) above-mentioned clear liquid 355L carries out pottery filter, and pottery filter sizes is 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, one-level
Membrane aperture 8000-10000 dalton, secondary membrane aperture 3000-5000 dalton, two-stage ultrafiltering, after terminating, use 50L purified water
Washing membranous system, obtained film dialysis clear liquid 400L, detected agmatine content 190.2g/L, yield 90.5%, crossed film dialysis clear liquid saturating
Light rate is more than 97%.
(4) take the above-mentioned film dialysis clear liquid 400L that crosses and be concentrated into the 40% of film dialysis clear liquid original volume, add 2kg activated carbon
Adsorption bleaching, after filtration, regulates pH to 6.0, and the ethanol 500L adding volume fraction 95% in gained feed liquid separates out crystal, mistake
Filter to obtain agmatine crude product, pure 69.3kg, yield 91.2%.
Embodiment 3
(1) using raw material is arginine, arginine decarboxylase (arginine decarboxylase is the arginine decarboxylase from antibacterial)
Lower bioenzymatic conversion feed liquid 800L containing agmatine produced of effect is placed in freeze dryer distributor pipe, carries out freeze thawing treatment, soon
Speed is cooled to-20 DEG C and is incubated 1 hour, is rapidly heated to 5 DEG C, is directly placed in centrifuge by the liquid in distributor pipe, and rejection filter is divided
Separate out part thalline；Liquid rejection filter gone out the most averagely imports in distributor pipe, fast cooling to-30 DEG C insulation 1 hour after again
It is warming up to 10 DEG C, is cooled to-35 DEG C in the same way and rises again again to 20 DEG C.
(2) the bioenzymatic conversion feed liquid (freeze thawing inactivation) that will rise again, adds the bioenzymatic conversion feed liquid quality 2.5% risen again
Flocculant, slowly put into tube centrifuge after stirring 1 hour, centrifugal rotational speed, at 12000～15000 revs/min, isolates bacterium
Body, continuous extraction clear liquid, obtain centrifugal clear liquid 760L.
(3) above-mentioned centrifugal clear liquid 760L carries out pottery filter, and pottery filter sizes is 20-100 μm；Ultrafiltration uses two-stage ultrafiltering,
One-level membrane aperture 8000-10000 dalton, secondary membrane aperture 3000-5000 dalton, two-stage ultrafiltering is after terminating, pure with 50L
Change water washing membranous system, obtained film dialysis clear liquid 805L, and detected agmatine content 184.34g/L, yield 92.3%, and crossed film dialysis
Clear liquid light transmittance is more than 97%.
(4) take the above-mentioned film dialysis clear liquid 400L that crosses and be concentrated into the 50% of film dialysis clear liquid original volume, add 2kg activated carbon
Adsorption bleaching, after filtration, regulates pH to 4.5, and the ethanol 500L adding volume fraction 95% in gained feed liquid separates out crystal, mistake
Filtering to obtain agmatine crude product, 60 DEG C of vacuum is dried, detects moisture 0.3%, obtains the pure 67.1kg of agmatine crude product, yield 91.0%.
Claims (10)
1. the method extracting agmatine, it is characterised in that: comprise the following steps:
(1) using raw material is arginine, the bioenzymatic conversion feed liquid containing agmatine produced under arginine decarboxylase effect
Carry out freeze thawing treatment, make arginine decarboxylase lose biological activity；
(2) take the bioenzymatic conversion feed liquid after inactivation and carry out high speed centrifugation, isolate thalline and protein matter, obtain centrifugal clear
Liquid；
(3) centrifugal clear liquid is crossed membranous system, remove thalline relic and macromolecular substances, obtained film dialysis clear liquid；
(4) by cross film dialysis clear liquid concentrate, after activated carbon decolorizing regulate pH carry out solvent crystal, filter agmatine is thick
Product.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that: in step (1), arginine decarboxylation
Enzyme is the arginine decarboxylase from antibacterial or plant-derived arginine decarboxylase.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that: in step (1), freeze thawing treatment,
Use-20 DEG C～5 DEG C ,-30 DEG C～the freeze thawing treatment scheme of 10 DEG C ,-35 DEG C～20 DEG C successively.
A kind of method extracting agmatine the most according to claim 3, it is characterised in that: in step (1), freeze thawing treatment,
The mode cutting out partial thalline of filtration is used after dissolving for the first time.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that: in step (2), after taking inactivation
After bioenzymatic conversion feed liquid adds chelating agent or flocculant, it is centrifuged.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that: in step (2), high speed centrifugation is adopted
With disk centrifugal separator or tube centrifuge, centrifugal rotational speed is at 12000～15000 revs/min.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that: in step (3), membranous system is pottery
Filter membrane and ultrafilter membrane.
A kind of method extracting agmatine the most according to claim 7, it is characterised in that: in step (3), filter sizes of making pottery
For 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, one-level membrane aperture 8000-10000 dalton, 3000-5000 road, secondary membrane aperture
Er Dun.
A kind of method extracting agmatine the most according to claim 1, it is characterised in that in step (4), pH is 3.0-
7.0。
A kind of method extracting agmatine the most according to claim 1, it is characterised in that in step (4), solvent is first
Alcohol, ethanol or isopropanol.
Priority Applications (1)
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| CN114058651A (en) * | 2021-09-30 | 2022-02-18 | 新泰市佳禾生物科技有限公司 | Preparation method of agmatine |
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| EP1260588A2 (en) * | 2001-05-25 | 2002-11-27 | Ajinomoto Co., Ltd. | Methods of producing agmatine and arginine decarboxylase |
| EP1270552A2 (en) * | 2001-05-25 | 2003-01-02 | Ajinomoto Co., Inc. | Production process for a guanidine derivative containing an amido group and for a salt thereof |
| CN104152478A (en) * | 2014-07-31 | 2014-11-19 | 洛阳华荣生物技术有限公司 | Method for coproducing D-arginine and gamatine through biotransformation |
| CN105861529A (en) * | 2016-05-27 | 2016-08-17 | 江南大学 | Arginine decarboxylase and application thereof |
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| EP1260588A2 (en) * | 2001-05-25 | 2002-11-27 | Ajinomoto Co., Ltd. | Methods of producing agmatine and arginine decarboxylase |
| EP1270552A2 (en) * | 2001-05-25 | 2003-01-02 | Ajinomoto Co., Inc. | Production process for a guanidine derivative containing an amido group and for a salt thereof |
| CN104152478A (en) * | 2014-07-31 | 2014-11-19 | 洛阳华荣生物技术有限公司 | Method for coproducing D-arginine and gamatine through biotransformation |
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| CN114058651A (en) * | 2021-09-30 | 2022-02-18 | 新泰市佳禾生物科技有限公司 | Preparation method of agmatine |
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Denomination of invention: A method for extracting agmatine Granted publication date: 20180601 Pledgee: Ningjin Sub branch of Xingtai Bank Co.,Ltd. Pledgor: JING JING PHARMACEUTICAL Co.,Ltd. Registration number: Y2024980043364 |