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Purification and characterization of the tyrosinase of Streptomyces michiganensis DSM 40015
Author(s) -
Philipp Stephan,
Held Thomas,
Kutzner Hans J.
Publication year - 1991
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.3620310412
Subject(s) - isoelectric point , chromatography , chemistry , sephadex , ammonium sulfate precipitation , size exclusion chromatography , isoelectric focusing , tyrosinase , enzyme , enzyme assay , substrate (aquarium) , hydrophilic interaction chromatography , electrophoresis , biochemistry , high performance liquid chromatography , biology , ecology
Tyrosinase of Streptomyces michiganensis DSM 40015 was purified 61‐fold from the culture broth: After a fractionated ammonium sulfate precipitation the enzyme was separated by hydrophobic interaction chromatography with Phenyl‐Sepharose and ionic interaction chromatography with CM‐Cellulose; finally a gel filtration with Sephadex G‐75 yielded 1.7% of the originally existent enzyme. SDS gel‐electrophoresis of the purified enzyme showed two bands representing a size of 34500 and 32000 dalton, respectively. However, by isoelectric focussing only one band could be found exhibiting an isoelectric point of approximately 9.0. Temperature and pH‐optimum of the enzyme activity were 33°C and pH 7.0, respectively. Whereas the enzyme is specific in regard to the aromatic part of its substrate variations of the aliphatic rest are tolerated.