SU532341A3 - Способ выделени фермента, катализирующего превращение креатинина в креатин и фермента, катализирующего превращение креатина в саркозин и мочевину - Google Patents
Способ выделени фермента, катализирующего превращение креатинина в креатин и фермента, катализирующего превращение креатина в саркозин и мочевинуInfo
- Publication number
- SU532341A3 SU532341A3 SU1781172A SU1781172A SU532341A3 SU 532341 A3 SU532341 A3 SU 532341A3 SU 1781172 A SU1781172 A SU 1781172A SU 1781172 A SU1781172 A SU 1781172A SU 532341 A3 SU532341 A3 SU 532341A3
- Authority
- SU
- USSR - Soviet Union
- Prior art keywords
- creatine
- conversion
- enzyme catalyzing
- creatinine
- sarcosine
- Prior art date
Links
- 238000000034 method Methods 0.000 title claims 2
- CVSVTCORWBXHQV-UHFFFAOYSA-N creatine Chemical compound NC(=[NH2+])N(C)CC([O-])=O CVSVTCORWBXHQV-UHFFFAOYSA-N 0.000 title description 8
- 229960003624 creatine Drugs 0.000 title description 4
- 239000006046 creatine Substances 0.000 title description 4
- 108090000790 Enzymes Proteins 0.000 title description 3
- 102000004190 Enzymes Human genes 0.000 title description 3
- 238000006243 chemical reaction Methods 0.000 title 2
- DDRJAANPRJIHGJ-UHFFFAOYSA-N creatinine Chemical compound CN1CC(=O)NC1=N DDRJAANPRJIHGJ-UHFFFAOYSA-N 0.000 title 2
- FSYKKLYZXJSNPZ-UHFFFAOYSA-N sarcosine Chemical compound C[NH2+]CC([O-])=O FSYKKLYZXJSNPZ-UHFFFAOYSA-N 0.000 title 2
- 108010077895 Sarcosine Proteins 0.000 title 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 title 1
- 239000004202 carbamide Substances 0.000 title 1
- 229940109239 creatinine Drugs 0.000 title 1
- 229940043230 sarcosine Drugs 0.000 title 1
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- 229920005654 Sephadex Polymers 0.000 claims description 4
- 239000012507 Sephadex™ Substances 0.000 claims description 4
- 239000002244 precipitate Substances 0.000 claims description 2
- 239000003960 organic solvent Substances 0.000 claims 1
- 239000000047 product Substances 0.000 claims 1
- 102000004169 proteins and genes Human genes 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 claims 1
- 108010066906 Creatininase Proteins 0.000 description 4
- 108010077078 Creatinase Proteins 0.000 description 3
- 239000008057 potassium phosphate buffer Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- LWIHDJKSTIGBAC-UHFFFAOYSA-K tripotassium phosphate Chemical compound [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 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
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical group N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 1
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 1
- 235000011130 ammonium sulphate Nutrition 0.000 description 1
- 239000003957 anion exchange resin Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000005342 ion exchange Methods 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 229910000160 potassium phosphate Inorganic materials 0.000 description 1
- 235000011009 potassium phosphates Nutrition 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
- C12N9/14—Hydrolases (3)
- C12N9/78—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5)
- C12N9/80—Hydrolases (3) acting on carbon to nitrogen bonds other than peptide bonds (3.5) acting on amide bonds in linear amides (3.5.1)
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Immunology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Enzymes And Modification Thereof (AREA)
Description
сить приблизительно в 2,5 раза при одинаковой удельной активюсти.
Получени как описано выше, креатинин-ами ,. имеет константу равновесн креатин 1,27 (37° С; рН 8,0).«феатинин
Константа Михаэлиса дл креашнина как субстрата KM 3,3-10М (37°С; рН8,0).
П р и м е р 2. Разделение креатинин-амидогидролазы и креатин- амидиногидролазы.
Выделение провод т, как в примере i. Однако, после злюировани креатинин-амидогидролазы;, с ионообменной колонки элюируют креатин-ами-. диногидролазу при помощи 0,2 М буферного раствора фосфата кали с рН8,О, содержащего 0,3 Nf NaCI (см. табл. 2).
Результаты активности и выхода препарата
П р и м е р 3. Первый этап выделени провод т, как в примере 1. 100 г осажденного изопропанолом а.1рца раствор ют в 100 мл 0,02 М буферного раствора . фосфата кали с рН 8,0 и смешивают с ДЕАЕ- Сефадексом, Ионообменную смолу отфильтровывают , промывают приблизительно 100 мл 0,08 М буферного раствора фосфата кали с рН 8,0 и дл общего злншровани обоих ферментов перемеишвают 15 мин при плюс 4° С со 100мл 0,2 М буферного раствора фосфата кали с рН 8,0, содержащего 0,3 М KCI и затем отфильтровьшают. В злюате наход тс оба фермента. Осаждением сульфатом аммони можно получать препарат
с 31,5 едмг креатинин-амидогидролазы и 1,1 едгм креатин-амидиногидролазы (см. табл. 3).
Таблица 1
2,8-1098,6
2,,2 2,16-10 32,6
1, 6,9
1,8-100,59
Разделение креатмнин-амидогидролазы и креатин-амидино гидролазы на анионообменной смоле ДЕАЕ-Сефадекс
После стадии осажде1ш изопропанолом
ДЕАЕ-Сефадекспромьшна вода
0,20 М, рН 8,0
(Элюаты)
0,50 М, рН 8,0
(Элюаты)
2,8
100
3,9
93
6,6
77
28,5
71
303
64
ТаСлица2
0,25
44
8604,9
27
3,0
41
0,13
Выделение креатинин-амидогидролазы и креатагт-амлднногидролазы из 100 г на сухой вес Alcaiigenes spec. WS 51400
Разрушение
Втора добавка изопропанола (осадок)
Claims (1)
- ДЕАЕ-Сефадекс элюат 0,5 М рН 8,0 Формула изобретени Способвьщелеии фермента,катал 1зиругацегопре вращение креатининав креатин, и фермента, катализирующего превращение креатина в саркозин и мочевину,.ТаблицаЗ2,4Ю6Ш0,0230,923,,2651,98-10 13,94,,1 1,16-10 31,5 из клеток Alcaiigenes spec WS 51400 или Penicilliurn WS 90001, отличающийс тем, что клетки разрушают, отдел ют балластш11е белки, осаждают активную фракшчю органическим растворителем, например изопропанолом, и выдел ют и очищают целевой продукт известными приемами.
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE2122298A DE2122298C3 (de) | 1971-05-05 | 1971-05-05 | Verfahren zur Gewinnung von Creatininamidohydrolase |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU532341A3 true SU532341A3 (ru) | 1976-10-15 |
Family
ID=5806965
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU1781172A SU532341A3 (ru) | 1971-05-05 | 1972-05-04 | Способ выделени фермента, катализирующего превращение креатинина в креатин и фермента, катализирующего превращение креатина в саркозин и мочевину |
Country Status (8)
| Country | Link |
|---|---|
| JP (1) | JPS5729150B1 (ru) |
| DK (1) | DK134026C (ru) |
| FI (1) | FI51358C (ru) |
| HU (1) | HU166364B (ru) |
| IL (1) | IL39362A (ru) |
| IT (1) | IT954975B (ru) |
| SE (1) | SE7900292L (ru) |
| SU (1) | SU532341A3 (ru) |
-
1972
- 1972-05-04 JP JP4453472A patent/JPS5729150B1/ja active Pending
- 1972-05-04 IT IT23890/72A patent/IT954975B/it active
- 1972-05-04 IL IL39362A patent/IL39362A/xx unknown
- 1972-05-04 SU SU1781172A patent/SU532341A3/ru active
- 1972-05-04 HU HUBO1369A patent/HU166364B/hu unknown
- 1972-05-04 FI FI721267A patent/FI51358C/fi active
- 1972-05-04 DK DK221372A patent/DK134026C/da not_active IP Right Cessation
-
1979
- 1979-01-12 SE SE7900292A patent/SE7900292L/xx unknown
Also Published As
| Publication number | Publication date |
|---|---|
| JPS5729150B1 (ru) | 1982-06-21 |
| FI51358B (ru) | 1976-08-31 |
| IL39362A (en) | 1974-11-29 |
| IT954975B (it) | 1973-09-15 |
| DK134026B (da) | 1976-08-30 |
| FI51358C (fi) | 1976-12-10 |
| SE7900292L (sv) | 1979-01-12 |
| DK134026C (da) | 1977-02-07 |
| HU166364B (ru) | 1975-03-28 |
| IL39362A0 (en) | 1972-07-26 |
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