Expression, purification and enzymatic characterization of Thermus thermophilus HB8 aspartate aminotransferase in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 278-283, 2007.
Artigo
em Chinês
| WPRIM
| ID: wpr-325379
ABSTRACT
To obtain thermostable aspartate aminotransferase, the gene aspC from an extremely thermophilic bacterium, Thermus thermophilus HB8 was cloned, and its product was overexpressed in Escherichia coli BL21 (DE3) and Rosetta (DE3). The expression in Rosetta (DP3) was more efficient. The optimum reactive pH was 7, and the recombinant enzyme activity changed little when incubated in the buffer of pH8 - 10 on 37 degrees C for 1 h. The optimum reactive temprature was 75 degrees C, and the recombinant enzyme was more stable on the temperature of 25 - 55 degrees C. The half life of recombinant enzyme on 65 degrees C was 3.5 h, on 75 degrees C was 2.5 h. KmKG was 7.559 mmol/L, VmaxKG was 0.086 mmol/(L x min), KmAsp was 2.031 mmol/L, VmaxAsp was 0.024 mmol/(L x min). Ca2+, Fe3+, Mn2+ inhibited enzyme activity softly.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Farmacologia
/
Aspartato Aminotransferases
/
Temperatura
/
Proteínas de Bactérias
/
Estabilidade Enzimática
/
Proteínas Recombinantes
/
Cinética
/
Regulação Bacteriana da Expressão Gênica
/
Regulação Enzimológica da Expressão Gênica
/
Thermus thermophilus
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2007
Tipo de documento:
Artigo
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