SU560614A1

SU560614A1 - The method of producing phospholipase d

Info

Publication number: SU560614A1
Application number: SU2324341A
Authority: SU
Inventors: Мирзаатхам Мирзахакимович Рахимов; Шухрат Раимджанович Мадьяров; Абдумумин Хакимович Абдумаликов
Original assignee: Среднеазиатский Научно-Исследовательский И Проектно-Конструкторский Институт Пищевой Промышленности
Priority date: 1976-02-11
Filing date: 1976-02-11
Publication date: 1977-06-05

Description

1one

Изобретение относитс к технической биохимии , а именно к способам получени фосфолипазы D и может быть использовано в медицине и-пищевой промышленности.The invention relates to technical biochemistry, in particular to methods for producing phospholipase D, and can be used in medicine and food industry.

Известен способ получени фосфолипазы D,включающий экстракцию белков из сем н хлопчатника, осаждение целевого фермента и его адсорбционную очистку из раствора 1.A known method for the preparation of phospholipase D involves the extraction of proteins from cotton seeds, the precipitation of the target enzyme and its adsorption purification from solution 1.

Одна полученна по этому способу фосфолипаза D нестабильна в растворах,содержащих сульфат аммони ,и тер ет свою каталитическую активность.One phospholipase D obtained by this method is unstable in solutions containing ammonium sulphate and loses its catalytic activity.

С целью повышени выхода и каталитической активности фермента по предлагаемому способу экстракцию белков осуществл ют трис-буфером с рН 7-9, при этом адсорбционную очис та у провод т на силикагеле.In order to increase the yield and catalytic activity of the enzyme according to the proposed method, the extraction of proteins is carried out with a Tris buffer with a pH of 7-9, while the adsorption peptide is carried out on silica gel.

Концентраци трис-буфера равна О,01-О ,1 М, а раствор имеет концентрацию 0,1-1 ,0 г/л. по белку.The concentration of Tris buffer is O, 01-O, 1 M, and the solution has a concentration of 0.1-1.0 g / l. by squirrel.

Способ заключаетс Б том, что экс1-ракцию белков, провод т трис-буфером концентрацией 0,01-0,1 М, рН 7-9 и адсорбируют фосфолипазу после растворени . Адсорбцию провод т из разбавленного раствора The method consists in the fact that the protein is extracted by a protein of 0.01-0.1 M, pH 7-9, and phospholipase is adsorbed after dissolution. The adsorption is carried out from a dilute solution.

конпентращ1ей 0,1-1,0 г/л по белку и кол- личестве силикагел 2О-120 г/п приcontent of 0.1-1.0 g / l for protein and amount of silica gel 2 O-120 g / p at

о,течение 15 мин - 24 ч.oh, for 15 minutes - 24 hours

Claims

Example, A portion of cotton cesiim is crushed and extracted with ten times O volume, 1 L Tris buffer, pH 8.8, HAM, and soluble proteins are separated from the precipitate by centrifugation. Two volumes of cooled acetone (-10 ° C) are added to the insufficient liquid and mixed. The precipitate formed is separated by decantation, washed with acetone and dissolved in water. The pH is adjusted to pH 6 with 1N. CIDCON and heat 15 mnn at 50 ° C. The precipitate is removed by centrifugation (15 min at 18000g). A solution of OD M is diluted with acetate buffer, pH 5.6 to concentrate O, 1 mg / m in protein content, and KSK K2 2 slicel is added to the solution at the rate of SO g / n and left to incubate phospholipase D is separated from the centrifugal / tem solution (1 O min EOOb). The use of the pressed method of phosphopipase D production increases the yield of fsjupolipase D 1.8-2.5 times; increases the catalytic activity of the drug popu- lar 3-6 times; fsyufolipaza D form in insoluble form adsorbed on silica gel and is reused. Formula of inventions 7-9, with adsorption purification being carried out on the silica gel.

2. A method according to claim 1, characterized in that the concentration of Tris buffer is 0.01-0.1 M, and the solution has a concentration of 0.1-1.0 g / l of protein. Sources of information taken into account during the examination: 1. Copyright certificate №353725, CL, A 61 K 35/78, 1972.