Purification and characterization of the tyrosinase of Streptomyces michiganensis DSM 40015

✍ Scribed by Stephan Philipp; Thomas Held; Prof. Dr. Hans J. Kutzner

• Publisher: John Wiley and Sons
• Year: 1991
• Tongue: English
• Weight: 493 KB
• Volume: 31
• Category: Article
• ISSN: 0233-111X
• DOI: 10.1002/jobm.3620310412

✦ Synopsis

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.