Physiological role of D‐aspartate oxidase in the assimilation and detoxification of D‐aspartate in the yeast Cryptococcus humicola

Abstract

The physiological role of D-aspartate oxidase (ChDASPO) in the yeast Cryptococcus humicola was analysed through the growth characteristics of a ChDASPO gene-disrupted strain (daspoDelta) and the expression profile of ChDASPO on various combinations of carbon and nitrogen sources. The daspoDelta strain, constructed by homologous integration of the yeast URA3 marker, grew as well as the wild-type strain on ammonium chloride, L-aspartate or D-alanine as the sole nitrogen source. In contrast, the daspoDelta strain did not grow at all on D-aspartate, not only as the sole nitrogen source but also as the sole carbon source or as the sole nitrogen and carbon source, and grew more slowly than the wild-type strain on D-glutamate as the sole nitrogen source. In the wild-type strain, the induction of ChDASPO activity strictly depended on the presence of D-aspartate and was little affected by the co-presence of ammonium chloride, but it was significantly reduced by the co-presence of both glucose and ammonium chloride, which, however, did not abolish the induction, allowing considerable expression of ChDASPO. This expression pattern was consistent with that shown by Northern blot analysis. The daspoDelta strain was more sensitive than the wild-type to the growth retardation by acidic D-amino acids, but not to that by the corresponding L-isomers or D-alanine. These results clearly show that in the yeast, DASPO plays an essential role in the assimilation of D-aspartate and acts as a detoxifying agent for D-aspartate.