RESUMEN

A gene of Aspergillus oryzae, ladA, which encodes L-arabinitol 4-dehydrogenase (EC 1.1.1.12), and its cDNA were cloned in Escherichia coli. The gene consisted of a 1209-bp coding region, interrupted by a 59-bp intron, which encoded a 382-amino-acid polypeptide (40,812 Da). The protein showed 67% identity to a well-studied L-arabinitol 4-dehydrogenase (Lad1) of Hypocrea jecorina. The cell-free extract of E. coli, which expressed ladA cDNA, showed L-arabinitol dehydrogenase activity with NAD+. It was also reactive for ribitol and xylitol.

Asunto(s)

Aspergillus oryzae/enzimología , Genes Fúngicos , NAD/metabolismo , Deshidrogenasas del Alcohol de Azúcar/genética , Deshidrogenasas del Alcohol de Azúcar/metabolismo , Secuencia de Aminoácidos , Aspergillus oryzae/genética , Clonación Molecular , Escherichia coli/genética , Exones/genética , Intrones/genética , Datos de Secuencia Molecular , Alineación de Secuencia

RESUMEN

XdhA, which encodes a xylitol dehydrogenase gene, was cloned from Aspergillus oryzae genomic DNA. It consists of 1214 bp structural region, which is interrupted by two introns, and encodes 358-amino-acid protein (38,197 Da). It is similar to the known NAD(+)-dependent xylitol dehydrogenase (EC 1.1.1.9). The gene was expressed in Escherichia coli BL21-AI using a T7 promoter. The cell-free extract of the transformant showed a 36.5 kDa band upon SDS-PAGE and NAD(+)-dependent xylitol dehydrogenase activity.