CN106278954B - A kind of method for extracting agmatine - Google Patents
A kind of method for extracting agmatine Download PDFInfo
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- CN106278954B CN106278954B CN201610684104.8A CN201610684104A CN106278954B CN 106278954 B CN106278954 B CN 106278954B CN 201610684104 A CN201610684104 A CN 201610684104A CN 106278954 B CN106278954 B CN 106278954B
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- QYPPJABKJHAVHS-UHFFFAOYSA-N Agmatine Natural products NCCCCNC(N)=N QYPPJABKJHAVHS-UHFFFAOYSA-N 0.000 title claims abstract description 35
- QYPPJABKJHAVHS-UHFFFAOYSA-P agmatinium(2+) Chemical compound NC(=[NH2+])NCCCC[NH3+] QYPPJABKJHAVHS-UHFFFAOYSA-P 0.000 title claims abstract description 35
- 238000000034 method Methods 0.000 title claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 64
- 238000006243 chemical reaction Methods 0.000 claims abstract description 22
- 238000000502 dialysis Methods 0.000 claims abstract description 22
- 102100032252 Antizyme inhibitor 2 Human genes 0.000 claims abstract description 20
- 101000798222 Homo sapiens Antizyme inhibitor 2 Proteins 0.000 claims abstract description 19
- 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
- 230000000694 effects Effects 0.000 claims abstract description 10
- 239000004475 Arginine Substances 0.000 claims abstract description 9
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims abstract description 9
- 239000002904 solvent Substances 0.000 claims abstract description 9
- 239000002994 raw material Substances 0.000 claims abstract description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 239000013078 crystal Substances 0.000 claims abstract description 6
- 238000000703 high-speed centrifugation Methods 0.000 claims abstract description 6
- 230000002779 inactivation Effects 0.000 claims abstract description 5
- 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 9
- 238000000108 ultra-filtration Methods 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 6
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 5
- 239000008139 complexing agent Substances 0.000 claims description 5
- 235000019441 ethanol Nutrition 0.000 claims description 5
- 238000001914 filtration Methods 0.000 claims description 5
- 102000004190 Enzymes Human genes 0.000 claims description 3
- 108090000790 Enzymes Proteins 0.000 claims description 3
- 238000006114 decarboxylation reaction Methods 0.000 claims description 3
- 238000005374 membrane filtration Methods 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims 1
- 235000014666 liquid concentrate Nutrition 0.000 claims 1
- 239000012043 crude product Substances 0.000 abstract description 9
- 239000000047 product Substances 0.000 abstract description 9
- 239000012535 impurity Substances 0.000 abstract description 6
- 238000012545 processing Methods 0.000 abstract description 4
- 239000007806 chemical reaction intermediate Substances 0.000 abstract description 3
- 150000001875 compounds Chemical class 0.000 abstract description 2
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 10
- 238000003786 synthesis reaction Methods 0.000 description 6
- 238000001816 cooling Methods 0.000 description 5
- 238000004321 preservation Methods 0.000 description 5
- 238000004061 bleaching Methods 0.000 description 4
- 238000001179 sorption measurement Methods 0.000 description 4
- 238000007086 side reaction Methods 0.000 description 3
- 238000003756 stirring Methods 0.000 description 3
- 238000010792 warming Methods 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 206010013786 Dry skin Diseases 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
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- RJKFOVLPORLFTN-LEKSSAKUSA-N Progesterone Chemical compound C1CC2=CC(=O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H](C(=O)C)[C@@]1(C)CC2 RJKFOVLPORLFTN-LEKSSAKUSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- HQABUPZFAYXKJW-UHFFFAOYSA-N butan-1-amine Chemical compound CCCCN HQABUPZFAYXKJW-UHFFFAOYSA-N 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 229940088598 enzyme Drugs 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
- UMGDCJDMYOKAJW-UHFFFAOYSA-N thiourea Chemical compound NC(N)=S UMGDCJDMYOKAJW-UHFFFAOYSA-N 0.000 description 2
- 238000002834 transmittance Methods 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 239000003643 water by type 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
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 208000007271 Substance Withdrawal Syndrome Diseases 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
- 239000002253 acid Substances 0.000 description 1
- 230000001154 acute effect Effects 0.000 description 1
- 238000007098 aminolysis reaction Methods 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
- 230000005540 biological transmission 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
- 210000004027 cell Anatomy 0.000 description 1
- 210000003169 central nervous system Anatomy 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 238000007596 consolidation process Methods 0.000 description 1
- 239000000431 corpus luteum hormone Substances 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- 229960001484 edetic acid Drugs 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
- 210000004051 gastric juice Anatomy 0.000 description 1
- 239000004519 grease Substances 0.000 description 1
- 239000000122 growth hormone Substances 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 210000003016 hypothalamus Anatomy 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 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
- 229940124636 opioid drug Drugs 0.000 description 1
- 229940111202 pepsin 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
- KOUKXHPPRFNWPP-UHFFFAOYSA-N pyrazine-2,5-dicarboxylic acid;hydrate Chemical compound O.OC(=O)C1=CN=C(C(O)=O)C=N1 KOUKXHPPRFNWPP-UHFFFAOYSA-N 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
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 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 methods for extracting agmatine, are related to the extractive technique field of compound.Comprise the following steps：（1）Raw material is used as arginine, the bioenzymatic conversion feed liquid containing agmatine generated under arginine decarboxylase effect carries out freeze thawing treatment, and arginine decarboxylase is made to lose bioactivity；（2) the bioenzymatic conversion feed liquid after inactivation is taken to carry out high speed centrifugation, thalline and protein matter is isolated, obtains centrifugal clear liquid；（3）Centrifugal clear liquid is crossed into membranous system, removes thalline relic and macromolecular substances, obtained film dialysis clear liquid；（4）Film dialysis clear liquid is concentrated, pH is adjusted after activated carbon decolorizing and carries out solvent crystal, filters to obtain agmatine crude product.This method, which can be handled quickly, obtains agmatine crude product, reduces chemical reaction intermediate processing steps, reduces impurity in product, simultaneously, capacity efficiency is high, at low cost, easy to operate, good product quality improves product quality and yield, creates good economic benefit.
Description
Technical field
The present invention relates to the extractive technique fields of compound.
Background technology
Agmatine（Gamatine）It is the substance of a multi-biological function, by the hormone for stimulating release hypothalamus
Carry out work, corpus luteum hormone and growth hormone can be promoted, agmatine there are multiple benefits, can improve body durability degree, to movement
Member and bodybuilders have certain benefit and can make its satisfaction, while can accelerate physical recovery, improve muscle mass, reduce fat
Grease improves body performance, and to pursuing, body is dynamic and helpful, guanidine of seeking each age level healthy lifestyles
Base butylamine can also improve nitric oxide production content by two kinds of approach, stimulate release nitric oxide and inhibit Nitric oxide syntheses
Synthesis（Nitric oxide syntheses can consume nitric oxide）.
There is data to show, in central nervous system, gamatine can enhance the analgesic effect of opioid drug, slow down drug resistance
Property generation and improve Acute Morphine withdrawal syndrome, in addition gamatine can also promote consolidation of memory.In cardiovascular system,
Gamatine can reduce heart rate, mean arterial pressure, left chamber pressure, peripheral vascular resistance and cardiac index etc., while gamatine is also
The secretion of hydrochloric acid in gastric juice and pepsin can be increased, make mucosal thickness thinning, deterioration stress caused by mucosa infection etc..There is research simultaneously
Show that agmatine also has the specific effect for inhibiting tumor cell proliferation, and it inhibits the effect of tumor cell proliferation with knurl
Cell line specificity.
At this stage, gamatine is mainly obtained by chemical synthesis in the market, primary raw material has Putriscine, O- first
Base isourea sulfate, sulfuric acid, alcohol etc., reaction process high energy consumption, yield is low, and impurity is more, and security risk is big, there is certain dirt to environment
Dye；Isosorbide-5-Nitrae-dibromobutane and potassium phthalimide are also used simultaneously as raw material, are substituted, aminolysis and guanidinated three step
The report of agmatine sulfate is made in reaction, this route reaction condition is mild, but side reaction is more, and cumbersome, and smell is difficult
It hears, reaction process use does not meet the production requirement of clean and effective compared with multi-solvent.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of method for extracting agmatine, this method can be handled quickly and obtained
Agmatine crude product is obtained, relatively traditional chemical synthesis effectively reduces chemical reaction intermediate processing steps, reduces intermediate
Side reaction and impurity in the process, the generation of give up solvent, exhaust gas, meanwhile, capacity efficiency is high, at low cost, easy to operate, product matter
It measures, reduces impurity in product, improve product quality and yield, create good economic benefit.
In order to solve the above technical problems, the technical solution used in the present invention is：A kind of method for extracting agmatine, including
Following steps：
（1）Raw material is used as arginine, in arginine decarboxylase（Arginine decarboxylase, ADC）Under effect
The bioenzymatic conversion feed liquid containing agmatine generated carries out freeze thawing treatment, and arginine decarboxylase is made to lose bioactivity；
（2) take inactivation after bioenzymatic conversion feed liquid carry out high speed centrifugation, isolate thalline and protein matter, obtain from
Heart clear liquid；
（3）Centrifugal clear liquid is crossed into membranous system, removes thalline relic and macromolecular substances, obtained film dialysis clear liquid；
（4）Film dialysis clear liquid is concentrated, pH is adjusted after activated carbon decolorizing and carries out solvent crystal, filters to obtain agmatine
Crude product.
Preferably, step（1）In, arginine decarboxylase is the arginine decarboxylase from bacterium or plant-derived smart ammonia
Acid decarboxylase.
Preferably, step（1）In, freeze thawing treatment uses -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 step（1）In, freeze thawing treatment isolates portion after being dissolved in first time by the way of filtering
Divide thalline.
Preferably, step（2）In, after the bioenzymatic conversion feed liquid after inactivation is taken to add in complexing agent or flocculant, carry out from
The heart.
It is further preferred that complexing agent used or flocculant have：Mercaptoethylmaine, polyacrylamide, thioacetic acid, thiocarbamide or
EDTA（Ethylenediamine tetra-acetic acid）.
Preferably, step（2）In, high speed centrifugation uses disk centrifugal separator or tube centrifuge, centrifugal rotational speed 12000~
15000 revs/min.
Preferably, step（2）In, high speed centrifugation uses stacked centrifuge, and centrifugal rotational speed is at 15000 revs/min.
Preferably, step（3）In, membranous system is pottery filter membrane and ultrafiltration membrane.
It is further preferred that step（3）In, pottery filter sizes are 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, level-one fenestra
Footpath 8000-10000 dalton, two level membrane aperture 3000-5000 dalton.
Step（3）In, ultrafiltration, two level membrane aperture or 1500-3000 dalton；
Film dialysis clear liquid was obtained after two-stage ultrafiltering, film is crossed and dialyses clear liquid light transmittance more than 97%.
Preferably, step（4）In, pH 3.0-7.0.
Preferably, step（4）In, will cross film dialysis clear liquid be concentrated into film dialysis clear liquid original volume 20-50% after, add
The medicinal carbon for entering agmatine quality 0.5%-3% in film dialysis clear liquid carries out adsorption bleaching.
Preferably, step（4）In, solvent is methanol, ethyl alcohol or isopropanol, and solvent dosage crosses film dialysis to adjust after pH
1.2-4.5 times of supernatant volume.
It is using advantageous effect caused by above-mentioned technical proposal：The method of the present invention can quickly handle acquisition agmatine
Crude product, relatively traditional chemical synthesis effectively reduce chemical reaction intermediate processing steps, reduce intermediate side reaction and
Impurity in the process, the generation of give up solvent, exhaust gas, meanwhile, capacity efficiency is high, and at low cost, easy to operate, good product quality subtracts
Lack impurity in product, improved product quality and yield, create good economic benefit.
Specific embodiment
The present invention will be further described in detail with reference to the specific embodiments.
Embodiment 1
（1）Raw material is used as arginine, in arginine decarboxylase（Arginine decarboxylase is the arginine decarboxylation from bacterium
Enzyme）The lower bioenzymatic conversion feed liquid 500L containing agmatine 105kg generated of effect is placed in freeze dryer distributor pipe, carries out freeze thawing
Processing, fast cooling to -20 DEG C heat preservation 1 it is small when, be rapidly heated to 5 DEG C, the liquid in distributor pipe be directly placed into centrifuge,
Rejection filter isolates part thalline；The liquid that rejection filter goes out averagely is imported in freeze dryer distributor pipe again, fast cooling to -30 DEG C of guarantors
10 DEG C are warming up to again after when temperature 1 is small, are cooled to -35 DEG C in the same way and are risen again again to 20 DEG C.
（2）The bioenzymatic conversion feed liquid that will be risen again（Freeze thawing inactivates), the complexing agent of addition bioenzymatic conversion feed liquid quality 2%,
Stir 1 it is small when after be slowly put into disk centrifugal separator, centrifugal rotational speed is isolated thalline, is continuously adopted at 12000~15000 revs/min
Go out clear liquid, obtain centrifugal clear liquid 470L.
（3）Above-mentioned clear liquid 470L is subjected to pottery filter, pottery filter sizes are 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, level-one
Membrane aperture 8000-10000 dalton, two level membrane aperture 3000-5000 dalton, two-stage ultrafiltering, after, with 50L purified waters
Membranous system is washed, obtained film dialysis clear liquid 500L, detects agmatine content 193g/L, yield 92.3% crosses film dialysis clear liquid light transmission
Rate is more than 97%.
（4）Take it is above-mentioned cross film dialyse clear liquid 500L be concentrated into film dialysis clear liquid original volume 50% after, add in 3kg activity
Charcoal adsorption bleaching, after filtering, adjusts pH to 5.0,500L isopropanols is added in into gained feed liquid, crystal is precipitated, filter to obtain agmatine
Crude product after 65 DEG C of dryings of vacuum, detects moisture 0.26%, pure 90.22kg, yield 93.5%.
Embodiment 2
（1）Raw material is used as arginine, in arginine decarboxylase（Arginine decarboxylase is from plant-derived arginine
Decarboxylase）The lower bioenzymatic conversion feed liquid containing agmatine generated of effect carries out freeze thawing treatment, loses arginine decarboxylase
Bioactivity.By freeze dryer distributor pipe, 80 are cooled to -20 DEG C, and 5L bioenzymatic conversion feed liquids are added in into every distributor pipe, permanent
When -20 DEG C of heat preservations 1 of temperature are small, it is rapidly heated to 5 DEG C, the liquid in distributor pipe is directly placed into centrifuge, rejection filter isolates portion
Thalline is divided to amount to 4.6kg；The liquid that rejection filter is gone out averagely is imported in distributor pipe again, fast cooling to -30 DEG C of heat preservations 1 it is small when after
10 DEG C are warming up to again, are cooled to -35 DEG C in the same way and are risen again again to 20 DEG C.
（2）The bioenzymatic conversion feed liquid that will be risen again（Freeze thawing inactivates), add in the bioenzymatic conversion feed liquid quality 3% risen again
Complexing agent, stirring 1 it is small when after be slowly put into disk centrifugal separator, centrifugal rotational speed isolates bacterium at 12000~15000 revs/min
Body continuously produces clear liquid, obtains centrifugal clear liquid 355L.
（3）Above-mentioned clear liquid 355L is subjected to pottery filter, pottery filter sizes are 20-100 μm；Ultrafiltration uses two-stage ultrafiltering, level-one
Membrane aperture 8000-10000 dalton, two level membrane aperture 3000-5000 dalton, two-stage ultrafiltering, after, with 50L purified waters
Membranous system is washed, obtained film dialysis clear liquid 400L, detects agmatine content 190.2g/L, it is saturating to cross film dialysis clear liquid for yield 90.5%
Light rate is more than 97%.
（4）The above-mentioned dialysis of film excessively clear liquid 400L is taken to be concentrated into the 40% of film dialysis clear liquid original volume, adds in 2kg activated carbons
Adsorption bleaching after filtering, adjusts pH to 6.0, and crystal, mistake is precipitated in the ethyl alcohol 500L that volume fraction 95% is added in into gained feed liquid
Filter to obtain agmatine crude product, pure 69.3kg, yield 91.2%.
Embodiment 3
（1）Raw material is used as arginine, in arginine decarboxylase（Arginine decarboxylase is the arginine decarboxylation from bacterium
Enzyme）The lower bioenzymatic conversion feed liquid 800L containing agmatine generated of effect is placed in freeze dryer distributor pipe, carries out freeze thawing treatment,
Fast cooling to -20 DEG C heat preservation 1 it is small when, be rapidly heated to 5 DEG C, the liquid in distributor pipe be directly placed into centrifuge, rejection filter
Isolate part thalline；The liquid that rejection filter is gone out averagely is imported in distributor pipe again, fast cooling to -30 DEG C of heat preservations 1 it is small when after
10 DEG C are warming up to again, are cooled to -35 DEG C in the same way and are risen again again to 20 DEG C.
（2）The bioenzymatic conversion feed liquid that will be risen again（Freeze thawing inactivates), add in the bioenzymatic conversion feed liquid quality 2.5% risen again
Flocculant, stirring 1 it is small when after be slowly put into tube centrifuge, centrifugal rotational speed isolates bacterium at 12000~15000 revs/min
Body continuously produces clear liquid, obtains centrifugal clear liquid 760L.
（3）Above-mentioned centrifugal clear liquid 760L is subjected to pottery filter, pottery filter sizes are 20-100 μm；Ultrafiltration uses two-stage ultrafiltering,
Level-one membrane aperture 8000-10000 dalton, two level membrane aperture 3000-5000 dalton, two-stage ultrafiltering, after, it is pure with 50L
Change water washing membranous system, obtained film dialysis clear liquid 805L, detect agmatine content 184.34g/L, yield 92.3% crosses film dialysis
Clear liquid light transmittance is more than 97%.
（4）The above-mentioned dialysis of film excessively clear liquid 400L is taken to be concentrated into the 50% of film dialysis clear liquid original volume, adds in 2kg activated carbons
Adsorption bleaching after filtering, adjusts pH to 4.5, and crystal, mistake is precipitated in the ethyl alcohol 500L that volume fraction 95% is added in into gained feed liquid
Agmatine crude product is filtered to obtain, after 60 DEG C of dryings of vacuum, moisture 0.3% is detected, obtains the pure 67.1kg of agmatine crude product, yield 91.0%.
Claims (7)
- A kind of 1. method for extracting agmatine, it is characterised in that：Comprise the following steps：(1) raw material is used as arginine, the bioenzymatic conversion feed liquid containing agmatine generated under arginine decarboxylase effect Freeze thawing treatment is carried out, arginine decarboxylase is made to lose bioactivity；(2) the bioenzymatic conversion feed liquid after inactivation is taken to carry out high speed centrifugation, isolates thalline and protein matter, it is clear to obtain centrifugation Liquid；(3) centrifugal clear liquid is crossed into membranous system, removes thalline relic and macromolecular substances, obtain film dialysis clear liquid；(4) by film dialysis clear liquid concentrate, after activated carbon decolorizing adjust pH carry out solvent crystal, filter agmatine is thick Product；In step (1), freeze thawing treatment uses -20 DEG C~5 DEG C, -30 DEG C~10 DEG C, -35 DEG C~20 DEG C of freeze thawing treatment side successively Case；In step (3), membranous system is pottery filter membrane and ultrafiltration membrane；Filter sizes of making pottery are 20-100 μm；Ultrafiltration use two-stage ultrafiltering, one Grade membrane aperture 8000-10000 dalton, two level membrane aperture 3000-5000 dalton.
- 2. a kind of method for extracting agmatine according to claim 1, it is characterised in that：In step (1), arginine decarboxylation Enzyme is the arginine decarboxylase from bacterium or plant-derived arginine decarboxylase.
- 3. a kind of method for extracting agmatine according to claim 1, it is characterised in that：In step (1), freeze thawing treatment, After being dissolved in first time part thalline is isolated by the way of filtering.
- 4. a kind of method for extracting agmatine according to claim 1, it is characterised in that：In step (2), after taking inactivation After bioenzymatic conversion feed liquid adds in complexing agent or flocculant, centrifuged.
- 5. a kind of method for extracting agmatine 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.
- 6. a kind of method for extracting agmatine according to claim 1, it is characterised in that in step (4), pH 3.0- 7.0。
- 7. it is according to claim 1 it is a kind of extract agmatine method, it is characterised in that in step (4), solvent for methanol, Ethyl alcohol or isopropanol.
Priority Applications (1)
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
Non-Patent Citations (1)
| Title |
|---|
| "Enzymatic production of agmatine by recombinant arginine decarboxylase";Sun Xia等;《Journal of Molecular Catalysis B: Enzymatic》;20150620;第121卷;第1-8页 * |
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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 |