SU698623A1 - Method of producing trypsin inhibitor - Google Patents

Method of producing trypsin inhibitor

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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
Application number
SU782620955A
Other languages
Russian (ru)
Inventor
Ицык Менделеевич Вайсблай
Original Assignee
Кишиневский Ордена Трудового Красного Знамени Государственный Университет Им. В.И.Ленина
Priority date (The priority date 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 date listed.)
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Application filed by Кишиневский Ордена Трудового Красного Знамени Государственный Университет Им. В.И.Ленина filed Critical Кишиневский Ордена Трудового Красного Знамени Государственный Университет Им. В.И.Ленина
Priority to SU782620955A priority Critical patent/SU698623A1/en
Application granted granted Critical
Publication of SU698623A1 publication Critical patent/SU698623A1/en

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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

SU782620955A 1978-05-22 1978-05-22 Method of producing trypsin inhibitor SU698623A1 (en)

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

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SU782620955A SU698623A1 (en) 1978-05-22 1978-05-22 Method of producing trypsin inhibitor

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Cited By (2)

* Cited by examiner, † Cited by third party
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
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

Cited By (3)

* Cited by examiner, † Cited by third party
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
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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