Molecular cloning and characterization of a N-acetylneuraminate lyase gene from Staphylococcus hominis / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 480-489, 2013.
Artigo
em Chinês
| WPRIM
| ID: wpr-233228
ABSTRACT
A N-acetylneuraminate lyase gene (shnal) from Staphylococcus hominis was cloned into pET-28a and expressed in Escherichia coli BL21 (DE3) host cells. The recombinant enzyme was purified and characterized. It is a homotetrameric enzyme with the optimum pH at 8.0 for the cleavage direction and the optimum pH and temperature were 7.5 and 45 degrees C for the synthetic direction. The activity of ShNAL is stable when incubated at 45 degrees C for 2 h but decreased rapidly over 50 degrees C. ShNAL showed high stability in a wide range pH from 5.0 to 10.0 with the residual activity being > 70% when the enzyme was incubated in different buffers at 4 degrees C for 24 h. Its K(m) towards N-acetylneuraminic acid, pyruvate and ManNAc were (4.0 +/- 0.2) mmol/L, (35.1 +/- 3.2) mmol/L and (131.7 +/- 12.1) mmol/L, respectively. The k(cat)/K(m) value of Neu5Ac, ManNAc, and Pyr for ShNAL were 1.9 L/(mmol x s), 0.08 L/(mmol x s) and 0.08 L/(mmol x s), respectively.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Temperatura
/
Proteínas de Bactérias
/
Estabilidade Enzimática
/
Proteínas Recombinantes
/
Clonagem Molecular
/
Staphylococcus hominis
/
Escherichia coli
/
Genética
/
Concentração de Íons de Hidrogênio
/
Oxo-Ácido-Liases
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2013
Tipo de documento:
Artigo
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