Molecular characterizations of two dehydroascorbate reductases from Selaginella moellendorffii / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 76-84, 2011.
Artigo
em Chinês
| WPRIM
| ID: wpr-351531
ABSTRACT
Plant dehydroascorbate reductase (DHAR) is a physiologically important reducing enzyme in the ascorbate-glutathione recycling reaction. In this study, two DHARs genes (SmDHAR1 and SmDHAR2) were isolated from Selaginella moellendorffii. The SmDHAR1 and SmDHAR2 genes encode two proteins of 218 and 241 amino acid residues, with a calculated molecular mass of 23.97 kDa and 27.33 kDa, respectively. The genomic sequence analysis showed SmDHAR1 and SmDHAR2 contained five and six introns, respectively. Reverse transcription PCR revealed that the SmDHAR1 and SmDHAR2 were constitutive expression genes in S. moellendorffii. The recombinant SmDHAR1 and SmDHAR2 proteins were overexpressed in E. coli, and were purified by Ni-affinity chromatography. The recombinant SmDHAR1 showed 116-fold higher enzymatic activity towards the substrate dehydroascorbate than recombinant SmDHAR2. The recombinant SmDHAR1 showed higher thermal stability than recombinant SmDHAR2. These results indicated obvious functional divergence between the duplicate genes SmDHAR1 and SmDHAR2.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Oxirredutases
/
Proteínas de Plantas
/
Proteínas Recombinantes
/
Dados de Sequência Molecular
/
Sequência de Bases
/
Química
/
Sequência de Aminoácidos
/
Clonagem Molecular
/
Análise de Sequência de DNA
/
DNA de Plantas
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2011
Tipo de documento:
Artigo
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