Purification and Characterization of a Liver-derived β-N-Acetylhexosaminidase from Marine Mammal Sotalia fluviatilis
| Autor: | J. E. Gomes Júnior, Thales L. Rocha, J. A. T. Melo, Robert N.G. Miller, R. M. Nascimento, Djair S.L. Souza, A. L. M. Lima, L. R. D. Abreu, Octávio Luis Franco, M. F. GROSSI-de-SÁ |
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| Rok vydání: | 2010 |
| Předmět: |
Dolphins
Carbohydrates chemistry.chemical_element Bioengineering Calcium Biochemistry Analytical Chemistry chemistry.chemical_compound Chitin biology.animal Animals Sodium dodecyl sulfate Chromatography Molecular mass biology Organic Chemistry Temperature Substrate (chemistry) Hydrogen-Ion Concentration beta-N-Acetylhexosaminidases Enzyme assay Sotalia fluviatilis Sodium selenate Kinetics Liver chemistry Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Chromatography Gel biology.protein Electrophoresis Polyacrylamide Gel |
| Zdroj: | The Protein Journal. 29:188-194 |
| ISSN: | 1875-8355 1572-3887 |
| DOI: | 10.1007/s10930-010-9239-3 |
| Popis: | A beta-N-Acetylhexosaminidase (EC 3.2.1.52) was purified from hepatic extracts of Sotalia fluviatilis, order Cetacea. The protein was purified by using ammonium sulfate fractionation and four subsequent chromatographies (Biogel A 1.5 m, Chitin, Deae-Biogel and hydroxyapatite resins). After these purification steps, the enzyme was purified 380.5-fold with an 8.4% yield. The molecular mass (10 kDa) was estimated by SDS-PAGE and MALDI-TOF analysis. A Km of 2.72 mM and Vmax 9.5 x 10(-6) micromol/(min x mg) were found for this enzyme, determined by p-nitrophenyl-beta-D: -hexosaminide substrate digestion. Optimal pH and temperature for beta-N-Acetylhexosaminidase activity were 5.0 and 60 degrees C, respectively. Enzyme activity was inhibited by sodium selenate (Na(2)SeO(4)), mercuric chloride (HgCl(2)) and sodium dodecyl sulfate (C(12)H(25)SO(4)Na), and activated by zinc, calcium, barium and lithium ions. Characterization of the beta-N-Acetylhexosaminidase in Sotalia fluviatilis can be a basis for physiological studies in this species. |
| Databáze: | OpenAIRE |
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