Purification and Properties of Thermostableβ-Tyrosinase from an Obligately Symbiotic Thermophile,Symbiobacterium thermophilum
| Autor: | Seibun Suzuki, Toshikatsu Hirahara, Jae-Kuk Shim, Sueharu Horinouchi, Teruhiko Beppu |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Gel electrophoresis
Chromatography biology Chemistry Thermophile Tyrosinase Organic Chemistry Size-exclusion chromatography Tyrosine phenol-lyase General Medicine Symbiobacterium thermophilum Applied Microbiology and Biotechnology Biochemistry Enzyme assay Analytical Chemistry Isoelectric point biology.protein Molecular Biology Biotechnology |
| Zdroj: | Bioscience, Biotechnology, and Biochemistry. 56:84-89 |
| ISSN: | 1347-6947 0916-8451 |
| DOI: | 10.1271/bbb.56.84 |
| Popis: | Thermostable β-tyrosinase (tyrosine phenol-lyase, E.C.4.1.99.2) was extracted from an obligately symbiotic and thermophilic bacterium, Symbiobacterium thermophilum, which grows only in co-cultivation with a specific thermophilic Bacillus sp., strain S. The enzyme was purified 300-fold to homogeneity with 4.2% recovery by ammonium sulfate fractionation and several steps of chromatography with anion-exchange, hydroxylapatite, and hydrophobic interaction columns. The enzyme has a molecular weight of approximately 200,000, as estimated by gel filtration column chromatography, and 48,000, as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, which indicates that the native enzyme is composed of four homologous subunits. The enzyme was stable up to 80°C and the optimum temperature for the activity was 80°C. The enzyme had an optimum pH at 7 and the isoelectric point of pH 4.8. The addition of K+ and NH4+ accelerated the enzyme activity. In contrast, Na+ and Mg2+ showed no effect. The enzyme ... |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|