Structural and functional analyses of a novel manganese-catalase from Bacillus subtilis R5.
Int J Biol Macromol
; 180: 222-233, 2021 Jun 01.
Article
en En
| MEDLINE
| ID: mdl-33737179
Catalases catalyze the decomposition of hydrogen peroxide into water and oxygen. Limited reports are available on characterization of manganese-catalases. We describe here molecular cloning and expression in Escherichia coli of a putative manganese-catalase gene from mesophilic bacterium, Bacillus subtilis R5. The gene product, CatBsu, produced as a soluble protein, was purified to apparent homogeneity and biochemically characterized. The absorption spectra and nonsignificant inhibition by sodium azide indicated that it is a manganese-catalase. The protein was in homohexameric form in solution, with a subunit molecular weight of 30 kDa, containing ~2 Mn2+ and ~1 Ca2+ per subunit. CatBsu showed highest activity at pH 8.0 and 55 °C. It was found to be highly active with a specific activity of 25,290 µmol min-1 mg-1 and apparent Km and kcat values of 98 mM and 1.27 × 104 s-1 subunit-1, respectively. Although from a mesophilic source, it exhibited a half-life of 2 h at 80 °C. Furthermore, the active site and metal binding residues in CatBsu were predicted by homology modelling and molecular docking. To the best of our knowledge, this is the first characterization of a manganese-catalase from genus Bacillus.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Bacillus subtilis
/
Proteínas Bacterianas
/
Catalasa
Tipo de estudio:
Prognostic_studies
Idioma:
En
Revista:
Int J Biol Macromol
Año:
2021
Tipo del documento:
Article
País de afiliación:
Pakistán
Pais de publicación:
Países Bajos