METHOD OF ENDONUCLEASE RESTRICTION BSE 21 I PREPARING

biotechnology, biochemistry, gene engineering, enzymes. SUBSTANCE: invention proposes a method of isolation of endonuclease restriction Bse 21 I recognizing and cleaving the nucleotide sequence 5'-GCTNAGG-3' that ensures to obtain enzyme no containing impurities of nonspecific nucleases an...

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Bibliographic Details
Main Authors KUVSHINOV V.N, USHAKOVA T.A, PUCHKOVA L.I, REPIN V.E, MIKHAJLOVA V.K
Format Patent
LanguageEnglish
Russian
Published 10.07.2002
Edition7
Subjects
BEER
BIOCHEMISTRY
CHEMISTRY
COMPOSITIONS THEREOF
CULTURE MEDIA
ENZYMOLOGY
METALLURGY
MICROBIOLOGY
MICROORGANISMS OR ENZYMES
MUTATION OR GENETIC ENGINEERING
PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS
SPIRITS
VINEGAR
WINE
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Summary:biotechnology, biochemistry, gene engineering, enzymes. SUBSTANCE: invention proposes a method of isolation of endonuclease restriction Bse 21 I recognizing and cleaving the nucleotide sequence 5'-GCTNAGG-3' that ensures to obtain enzyme no containing impurities of nonspecific nucleases and phosphatases and with minimal amount of protein. The yield of enzyme is 3 000 000 U per 10 g of wetted biomass. Isolation of endonuclease restriction Bse 21 I is carried out at the room temperature and pH = 7.5 by fractionation of cellular homogenate in two-phase system polyethylene glycol/dextran in the presence of NaCl taken in concentration 0.2-0.3 M. Purification of enzyme is carried out by chromatography on phosphocellulose P-11 and hydroxylapatite successively. Method provides to obtain up to 30 000 U of enzyme activity/g of biomass that increases the yield of product and decreases the technological process of isolation to 40 h. EFFECT: improved method of preparing, increased yield of enzyme. 1 dwg, 1 tbl, 5 ex
Bibliography:Application Number: RU19990107352