Data/hora: 08/06/2024 15:52:25

Provedor de dados: | 74
País: | Brazil
Título: | Cloning, expression and characterization of alcohol dehydrogenases in the silkworm Bombyx mori
Autores: | Wang,Nan; Shi,Haifeng; Yao,Qin; Zhou,Yang; Kang,Lequn; Chen,Huiqin; Chen,Keping.
Data: | 2011-01-01
Ano: | 2011
Palavras-chave: | 5'-RACE PCRADH; Enzymatic activity; Recombinant protein.
Resumo: | Alcohol dehydrogenases (ADH) are a class of enzymes that catalyze the reversible oxidation of alcohols to corresponding aldehydes or ketones, by using either nicotinamide adenine dinucleotide (NAD) or nicotinamide adenine dinucleotide phosphate (NADP), as coenzymes. In this study, a short-chain ADH gene was identified in Bombyx mori by 5'-RACE PCR. This is the first time the coding region of BmADH has been cloned, expressed, purified and then characterized. The cDNA fragment encoding the BmADH protein was amplified from a pool of silkworm cDNAs by PCR, and then cloned into E. coli expression vector pET-30a(+). The recombinant His-tagged BmADH protein was expressed in E. coli BL21 (DE3), and then purified by metal chelating affinity chromatography. The soluble recombinant BmADH, produced at low-growth temperature, was instrumental in catalyzing the ethanol-dependent reduction of NAD+, thereby indicating ethanol as one of the substrates of BmADH.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200013
Editor: | Sociedade Brasileira de Genética
Relação: | 10.1590/S1415-47572011000200013
Formato: | text/html
Fonte: | Genetics and Molecular Biology  v.34 n.2 2011
Fonte: | info:eu-repo/semantics/openAccess