JPS6154398B2 - - Google Patents
Info
- Publication number
- JPS6154398B2 JPS6154398B2 JP53014549A JP1454978A JPS6154398B2 JP S6154398 B2 JPS6154398 B2 JP S6154398B2 JP 53014549 A JP53014549 A JP 53014549A JP 1454978 A JP1454978 A JP 1454978A JP S6154398 B2 JPS6154398 B2 JP S6154398B2
- Authority
- JP
- Japan
- Prior art keywords
- heparin
- heparitinase
- agarose
- solution
- immobilized
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- 108010083213 heparitinsulfate lyase Proteins 0.000 claims description 55
- 229920000669 heparin Polymers 0.000 claims description 42
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 claims description 41
- 229960002897 heparin Drugs 0.000 claims description 41
- 229920000936 Agarose Polymers 0.000 claims description 36
- 238000004440 column chromatography Methods 0.000 claims description 8
- 239000000284 extract Substances 0.000 claims description 8
- 229910052588 hydroxylapatite Inorganic materials 0.000 claims description 8
- XYJRXVWERLGGKC-UHFFFAOYSA-D pentacalcium;hydroxide;triphosphate Chemical compound [OH-].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O XYJRXVWERLGGKC-UHFFFAOYSA-D 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 7
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 6
- 102000007327 Protamines Human genes 0.000 claims description 5
- 108010007568 Protamines Proteins 0.000 claims description 5
- 241000589565 Flavobacterium Species 0.000 claims description 4
- 239000003795 chemical substances by application Substances 0.000 claims description 4
- 229940048914 protamine Drugs 0.000 claims description 3
- 230000000813 microbial effect Effects 0.000 claims 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 18
- 239000000243 solution Substances 0.000 description 18
- 210000004027 cell Anatomy 0.000 description 13
- 230000001580 bacterial effect Effects 0.000 description 12
- 238000002360 preparation method Methods 0.000 description 12
- 102000004190 Enzymes Human genes 0.000 description 11
- 108090000790 Enzymes Proteins 0.000 description 11
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 10
- 238000000034 method Methods 0.000 description 10
- 238000000746 purification Methods 0.000 description 10
- 239000000872 buffer Substances 0.000 description 9
- 230000000694 effects Effects 0.000 description 9
- 230000001965 increasing effect Effects 0.000 description 9
- 239000011780 sodium chloride Substances 0.000 description 9
- 238000010828 elution Methods 0.000 description 7
- 230000000052 comparative effect Effects 0.000 description 6
- 101710106625 Chondroitinase-AC Proteins 0.000 description 5
- 108010048429 chondroitinase B Proteins 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 239000008363 phosphate buffer Substances 0.000 description 5
- 241000605114 Pedobacter heparinus Species 0.000 description 4
- 229920002684 Sepharose Polymers 0.000 description 4
- 230000002378 acidificating effect Effects 0.000 description 4
- 239000007853 buffer solution Substances 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 238000011084 recovery Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000007796 conventional method Methods 0.000 description 3
- AVJBPWGFOQAPRH-FWMKGIEWSA-L dermatan sulfate Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@H](OS([O-])(=O)=O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](C([O-])=O)O1 AVJBPWGFOQAPRH-FWMKGIEWSA-L 0.000 description 3
- 125000000524 functional group Chemical group 0.000 description 3
- 230000003100 immobilizing effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide Chemical compound CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 description 2
- SQDAZGGFXASXDW-UHFFFAOYSA-N 5-bromo-2-(trifluoromethoxy)pyridine Chemical compound FC(F)(F)OC1=CC=C(Br)C=N1 SQDAZGGFXASXDW-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 229920001287 Chondroitin sulfate Polymers 0.000 description 2
- 229920002683 Glycosaminoglycan Polymers 0.000 description 2
- 108010022901 Heparin Lyase Proteins 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 210000004204 blood vessel Anatomy 0.000 description 2
- 229940059329 chondroitin sulfate Drugs 0.000 description 2
- ATDGTVJJHBUTRL-UHFFFAOYSA-N cyanogen bromide Chemical compound BrC#N ATDGTVJJHBUTRL-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000005194 fractionation Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 229950008679 protamine sulfate Drugs 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- -1 For example Polymers 0.000 description 1
- 108010023736 Chondroitinases and Chondroitin Lyases Proteins 0.000 description 1
- 102000011413 Chondroitinases and Chondroitin Lyases Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000008351 acetate buffer Substances 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 239000011324 bead Substances 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000023555 blood coagulation Effects 0.000 description 1
- VSGNNIFQASZAOI-UHFFFAOYSA-L calcium acetate Chemical compound [Ca+2].CC([O-])=O.CC([O-])=O VSGNNIFQASZAOI-UHFFFAOYSA-L 0.000 description 1
- 239000001639 calcium acetate Substances 0.000 description 1
- 229960005147 calcium acetate Drugs 0.000 description 1
- 235000011092 calcium acetate Nutrition 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 150000001718 carbodiimides Chemical class 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000019522 cellular metabolic process Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000000593 degrading effect Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 125000003700 epoxy group Chemical group 0.000 description 1
- 238000002523 gelfiltration Methods 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- ZFGMDIBRIDKWMY-PASTXAENSA-N heparin Chemical compound CC(O)=N[C@@H]1[C@@H](O)[C@H](O)[C@@H](COS(O)(=O)=O)O[C@@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O[C@H]2[C@@H]([C@@H](OS(O)(=O)=O)[C@@H](O[C@@H]3[C@@H](OC(O)[C@H](OS(O)(=O)=O)[C@H]3O)C(O)=O)O[C@@H]2O)CS(O)(=O)=O)[C@H](O)[C@H]1O ZFGMDIBRIDKWMY-PASTXAENSA-N 0.000 description 1
- 229960001008 heparin sodium Drugs 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000000704 physical effect Effects 0.000 description 1
- 230000008288 physiological mechanism Effects 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000002525 ultrasonication Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Landscapes
- Enzymes And Modification Thereof (AREA)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1454978A JPS54107585A (en) | 1978-02-10 | 1978-02-10 | Preparation of pure heparitinase |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP1454978A JPS54107585A (en) | 1978-02-10 | 1978-02-10 | Preparation of pure heparitinase |
Publications (2)
Publication Number | Publication Date |
---|---|
JPS54107585A JPS54107585A (en) | 1979-08-23 |
JPS6154398B2 true JPS6154398B2 (enrdf_load_stackoverflow) | 1986-11-21 |
Family
ID=11864224
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP1454978A Granted JPS54107585A (en) | 1978-02-10 | 1978-02-10 | Preparation of pure heparitinase |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPS54107585A (enrdf_load_stackoverflow) |
-
1978
- 1978-02-10 JP JP1454978A patent/JPS54107585A/ja active Granted
Also Published As
Publication number | Publication date |
---|---|
JPS54107585A (en) | 1979-08-23 |
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