Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 1 de 1
Filtrar
Añadir filtros








Intervalo de año
1.
Chinese Journal of Biotechnology ; (12): 1108-1111, 2008.
Artículo en Chino | WPRIM | ID: wpr-342784

RESUMEN

The xylose reductase of Pichia stipitis is one of the most important enzymes. It can be used to build up recombinant Saccharomyces cerevisiae strain for utilizing xylose and producing ethanol. Intercellular redox imbalance caused by NADPH preference over NADH for Pichia stipitis xylose reductase (PsXR) has been considered to be one of the main factors for poor ethanol productivity. Some key amino acids of PsXR, which affect the activity or coenzyme preference, were investigated in our previous study. In this study, Lys21 were rational designed for site-directed mutagenesis to alter coenzyme specificity of PsXR from NADPH and NADH into single NADH. The wild gene and mutagenesis genes were ligated into pET28b, and were transferred into E.coli BL21(DE3). After induced by IPTG, the xylose reductases were purified. Purified mutants K21A (Lys21-->Ala), K21R(Lys21-->Arg) were characterized by steady-state kinetic analysis. The results showed that the coenzyme dependence of K21A was completely reversed to NADH.


Asunto(s)
Aldehído Reductasa , Metabolismo , Sustitución de Aminoácidos , Genética , Coenzimas , Farmacología , Escherichia coli , Genética , Metabolismo , Etanol , Farmacología , Lisina , Genética , Mutagénesis Sitio-Dirigida , NAD , Metabolismo , NADP , Metabolismo , Pichia , Química , Genética , Metabolismo , Proteínas Recombinantes , Genética , Metabolismo , Recombinación Genética , Xilosa , Farmacología
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA