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Partial purification and characterization of anhydrotetracycline oxygenase of Streptomyces aureofaciens
Author(s) -
Vančurová Ivana,
Flieger Miroslav,
Volc Jindřich,
Beneš Milan J.,
Novotná Jana,
Neužil Jiří,
Běhal Vladislav
Publication year - 1987
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.3620270915
Subject(s) - isoelectric point , sephadex , chromatography , size exclusion chromatography , chemistry , isoelectric focusing , streptomyces aureofaciens , oxygenase , enzyme , hydrophilic interaction chromatography , thiol , biochemistry , high performance liquid chromatography , streptomyces , biology , bacteria , genetics
Anhydrotetracycline oxygenase was purified both by affinity chromatography and by hydrophobic interaction chromatography. Molecular weight of anhydrotetracycline oxygenase was determined to be 115,000 by Sephadex G‐200 gel filtration. Using preparative isoelectric focusing the isoelectric point of the enzyme was estimated to be 5.3. The enzyme showed a sensitivity to thiol‐specific inhibitors. During the hydrophobic interaction purification step, the activity dropped considerably. Reactivation occurred when a heat treated crude extract was added to the reaction mixture.