Characterization of xylitol dehydrogenase from debaryomyces hansenii

Abstract

The xylitol dehydrogenase (EC 1.1.1.9) from xylose-grown cells ofDebaryomyces hansenii was partially purified in two Chromatographic steps, and characterization studies were carried out in order to inves tigate the role of the xylitol dehydrogenase-catalyzed step in the regu lation of D-xylose metabolism. The enzyme was most active at pH 9.0–9.5, and exhibited a broad polyol specificity. The Michaelis con stants for xylitol and NAD+ were 16.5 and 0.55 mM, respectively. Ca2+, Mg2+, and Mn2+ did not affect the enzyme activity. Conversely, Zn2+, Cd2+, and Co2+ strongly inhibited the enzyme activity. It was concluded that NAD+-xylitol dehydrogenase from D.hansenii has similarities with other xylose-fermenting yeasts in respect to optimal pH, substrate specificity, and Km value for xylitol, and therefore should be named L-iditol:NAD+-5-oxidoreductase (EC 1.1.1.14). The reason D.hansenii is a good xylitol producer is not because of its value of Km for xylitol, which is low enough to assure its fast oxidation by NAD+ xylitol dehydrogenase. However, a higher Km value of xylitol dehydro genase for NAD+ compared to theK m values of other xylose-ferment ing yeasts may be responsible for the higher xylitol yields.