SU698623A1 - Method of producing trypsin inhibitor - Google Patents
Method of producing trypsin inhibitorInfo
- Publication number
- SU698623A1 SU698623A1 SU782620955A SU2620955A SU698623A1 SU 698623 A1 SU698623 A1 SU 698623A1 SU 782620955 A SU782620955 A SU 782620955A SU 2620955 A SU2620955 A SU 2620955A SU 698623 A1 SU698623 A1 SU 698623A1
- Authority
- SU
- USSR - Soviet Union
- Prior art keywords
- trypsin inhibitor
- producing
- producing trypsin
- distilled water
- inhibitor
- Prior art date
Links
- 238000000034 method Methods 0.000 title description 5
- 101710162629 Trypsin inhibitor Proteins 0.000 title description 4
- 229940122618 Trypsin inhibitor Drugs 0.000 title description 4
- 239000002753 trypsin inhibitor Substances 0.000 title description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 235000010582 Pisum sativum Nutrition 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- JVIPLYCGEZUBIO-UHFFFAOYSA-N 2-(4-fluorophenyl)-1,3-dioxoisoindole-5-carboxylic acid Chemical compound O=C1C2=CC(C(=O)O)=CC=C2C(=O)N1C1=CC=C(F)C=C1 JVIPLYCGEZUBIO-UHFFFAOYSA-N 0.000 description 2
- 229920001425 Diethylaminoethyl cellulose Polymers 0.000 description 2
- 240000004713 Pisum sativum Species 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 description 2
- 235000011130 ammonium sulphate Nutrition 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 235000013312 flour Nutrition 0.000 description 2
- 239000012465 retentate Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 229920000298 Cellophane Polymers 0.000 description 1
- 241000219843 Pisum Species 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 239000006174 pH buffer Substances 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000011347 resin Substances 0.000 description 1
- 229920005989 resin Polymers 0.000 description 1
- 238000005185 salting out Methods 0.000 description 1
- 239000013049 sediment Substances 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Landscapes
- Peptides Or Proteins (AREA)
Description
(54) СПОСОБ ПОЛУЧЕНИЯ ИНГИБИТОРА ТРИПСИНА(54) METHOD FOR OBTAINING TRIPSIN INHIBITOR
II
Изобретение относитс к химико-фармацевтической промышленности и касаетс получени ингибитора трипсина.This invention relates to the chemical and pharmaceutical industry and relates to the preparation of a trypsin inhibitor.
Известен способ получени ингибитора трипсина из сем н гороха путем экстракции раствором серной кислоты, подщелачивани до рН 7, 6, высаливани сульфатом аммони от 20 до 80% насыщени , хроматографии на трипстювой смоле диализом и высушивани 1.A method is known for obtaining a trypsin inhibitor from pea seeds by extracting with a solution of sulfuric acid, basifying to pH 7,6, salting out ammonium sulfate from 20 to 80% saturation, chromatography on triptych resin by dialysis and drying 1.
Однако этот способ не обеспечивает высокого выхода ингибитора трипсина (из 150 г муки - 1,2 мг).However, this method does not provide a high yield of trypsin inhibitor (from 150 g of flour - 1.2 mg).
Целью изобретени вл етс увеличение выхода целевого продукта.The aim of the invention is to increase the yield of the target product.
Эта цель достигаетс тем, что экстрагирование провод т из сем н гороха абиссинского дистиллированной водой, экстракт подкисл ют до рН 4,0-4,3 и хроматографиругот на диэтиламиноэтил целлюлозе при рН 8,15-8,28.This goal is achieved in that the extraction is carried out from the seeds of Abyssinian peas with distilled water, the extract is acidified to a pH of 4.0-4.3, and chromatography on diethylaminoethyl cellulose at a pH of 8.15-8.28.
На чертеже приведена хроматограмма, по сн юща предлагаемый способ.The drawing shows a chromatogram explaining the proposed method.
Пример. 150г гороха абиссинского экстрагируют 1,5 л дистиллированной воды в течение 1 ч при переме1пивании. ПолученнуюExample. 150 g of Abyssinian peas are extracted with 1.5 liters of distilled water for 1 hour while stirring. Received
суспензию центрифугируют 30 мин при 5500 д. Экстракт подкисл ют 0,5 ННС до рН 4,0 и снова таким же образом центрифугируют.the suspension is centrifuged for 30 minutes at 5500 d. The extract is acidified with 0.5 HNS to pH 4.0 and centrifuged again in the same manner.
Из полученной надосадочной жидкости высаливают сульфатом аммони фракцию 0-75 .Осадок суспендируют в 29-53 мл 0,1 н IMaCl, рН 8,20, внос т в цегтофановые мешки диаметром 2,0-2,5 см и диализчтот ; против дистиллированной воды рН 8,2 до отрицателыюй реакции на сульфат-ион. Ретентат (содержимое целлофановых мешков) центрифугируют, осадок отбрасывают , а надосадочн то жидкость нанос т на колонку с диэтиламинозтил-целлюлозой (32 х X 40 см), уравновешенной предварительно трис-НС1 буфером 0,025 М, рН 8,20 с ионной силой 0,0135.The resulting supernatant was salted out with ammonium sulfate fraction 0-75. The sediment was suspended in 29-53 ml of 0.1 N IMaCl, pH 8.20, introduced into cegtophan bags with a diameter of 2.0-2.5 cm and dialyztot; against distilled water pH 8.2 to a negative reaction to sulfate ion. The retentate (the contents of cellophane bags) is centrifuged, the precipitate is discarded, and the supernatant is applied to a column with diethylaminoethyl-cellulose (32 x X 40 cm), pre-equilibrated with Tris-HC1 buffer of 0.025 M, pH 8.20 with ionic strength of 0.0135 .
Белки элюируют этим же буфером на коллекторе . На полученной хроматограмме видно, что ингибирует трипсин только белок второго пика. Этот белок высаливают, диализуют и ретентат лиофилизуют Все операции с белками провод т на холоде (4-5°С), а хроматографнюпри комнатной температуре (17-20°С). Из указанного выше количества муки при рН буфераProteins are eluted with the same buffer on the collector. The resulting chromatogram shows that trypsin inhibits only the protein of the second peak. This protein is salted out, dialyzed and the retentate is lyophilized. All operations with proteins are carried out in the cold (4-5 ° C), and chromatographically at room temperature (17-20 ° C). From the above amount of flour at pH buffer
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU782620955A SU698623A1 (en) | 1978-05-22 | 1978-05-22 | Method of producing trypsin inhibitor |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SU782620955A SU698623A1 (en) | 1978-05-22 | 1978-05-22 | Method of producing trypsin inhibitor |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| SU698623A1 true SU698623A1 (en) | 1979-11-25 |
Family
ID=20766880
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| SU782620955A SU698623A1 (en) | 1978-05-22 | 1978-05-22 | Method of producing trypsin inhibitor |
Country Status (1)
| Country | Link |
|---|---|
| SU (1) | SU698623A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998033388A1 (en) * | 1997-01-29 | 1998-08-06 | Her Majesty In Right Of Canada As Represented By Agriculture And Agri-Food Canada | Insecticidal factor from field peas |
| RU2197252C1 (en) * | 2001-09-04 | 2003-01-27 | Общество с ограниченной ответственностью "НОМЕД" | Method of preparation "ingiprol" preparing |
| RU2524668C1 (en) * | 2013-05-06 | 2014-07-27 | Государственное бюджетное образовательное учреждение высшего профессионального образования "Астраханская государственная медицинская академия" Министерства здравоохранения Российской Федерации (ГБОУ ВПО АГМА Минздрава России) | Method for recovery of sperm-specific human antitrypsin |
-
1978
- 1978-05-22 SU SU782620955A patent/SU698623A1/en active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1998033388A1 (en) * | 1997-01-29 | 1998-08-06 | Her Majesty In Right Of Canada As Represented By Agriculture And Agri-Food Canada | Insecticidal factor from field peas |
| US5955082A (en) * | 1997-01-29 | 1999-09-21 | Her Majesty The Queen In Right Of Canada, As Represented By Agriculture And Agri-Food Canada | Insecticidal factor from field peas |
| RU2197252C1 (en) * | 2001-09-04 | 2003-01-27 | Общество с ограниченной ответственностью "НОМЕД" | Method of preparation "ingiprol" preparing |
| RU2524668C1 (en) * | 2013-05-06 | 2014-07-27 | Государственное бюджетное образовательное учреждение высшего профессионального образования "Астраханская государственная медицинская академия" Министерства здравоохранения Российской Федерации (ГБОУ ВПО АГМА Минздрава России) | Method for recovery of sperm-specific human antitrypsin |
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