METHOD FOR PRODUCING PHAGUE SPZ7 ENZYMES Russian patent published in 2012 - IPC A61K39/00 A61P31/04 C12N9/00 

Abstract RU 2460537 C2

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 Hottinger's 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.

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RU 2 460 537 C2

Authors

Levashov Pavel Andreevich

Popov Denis Viktorovich

Popova Valentina Mikhajlovna

Djatlov Ivan Alekseevich

Zhilenkov Evgenij Leonidovich

Morozova Ol'Ga Alekseevna

Belogurova Natal'Ja Georgievna

Pugachev Vladimir Georgievich

Repin Vladimir Evgen'Evich

Kljachko Natal'Ja L'Vovna

Levashov Andrej Vadimovich

Sedov Sergej Alekseevich

Legotskij Sergej Aleksandrovich

Dates

2012-09-10Published

2009-03-31Filed