LEVASHOV PAVEL ANDREEVICH,Левашов Павел Андреевич (RU),POPOV DENIS VIKTOROVICH,Попов Денис Викторович (RU),POPOVA VALENTINA MIKHAJLOVNA,Попова Валентина Михайловна (RU),DJATLOV IVAN ALEKSEEVICH,Дятлов
申请号：
RU2009111537/10
公开号：
RU0002460537C2
申请日:
2009.03.31
申请国别(地区):
RU
年份:
2012
代理人:
摘要：
FIELD: medicine.SUBSTANCE: invention relates to biochemistry. A method for producing phague SPZ7 enzymes from the collection of Federal State Institution State Scientific Centre of Applied Microbiology and Biotechnology implies as follows: the culture Salmonella enteritidis is introduced into Hottingers broth and grown. The grown culture is added with the purified phagolysate SPZ7 and cultured with aeration. The Salmonella enteritidis cells are deposited in a centrifuge. The produced cells are placed in tris-HCl buffer mixture 0.05 M of pH 7.0 containing ribonuclease, and cultured. It is added with ethylenediaminetetraacetate to the concentration of 5 mM the prepared suspension is centrifuged the supernatant is separated from the residue and fultered. Then the enzyme is deposited by adding (NH4)2SO4 to 100% saturation at temperature 0°C. Residual protein is dissolved in the buffer mixture the prepared enzyme solution is introduced in a chromatographic column with modified dextrane balanced by the buffer mixture tris-HCl 0.02 M pH=7.5 containing NaCl 0.15 M and ethylenediaminetetraacetate 1 mM chromatographic separation is performed at eluent feed rate 24 ml/h the prepared enzyme solution is further purified by ion-exchange chromatography. Chromatographic separation is performed at temperature 2-3°C at eluent feed rate 35 ml/h fractions containing 70% of total enzyme activity are separated and combined.EFFECT: invention enables avoiding many contra-indications and side effects observed when using live phague preparations.Изобретение относится к биохимии. Способ получения ферментов фага SPZ7 из коллекции федерального государственного научного учреждения "Государственный научный центр прикладной микробиологии и биотехнологии", заключается в следующем. В бульон Хоттингера вносят культуру Salmonella enteritidis и подращивают ее. В полученную культуру добавляют очищенный фаголизат SPZ7 и культивируют с аэрацией. Осаждают клетки Salmonella enteritidis на центрифуге. Получе
