CLONING OF THE ASPERGILLUS-NIGER GENE ENCODING ALPHA-L-ARABINOFURANOSIDASE-A

Using L-arabitol as an inducer, simple induction conditions were established that resulted in high-level expression of alpha-L-arabinofuranosidase A by an Aspergillus niger D-xylulose kinase mutant strain. These conditions were adapted to construct a cDNA expression library from which an alpha-L-arabinofuranosidase A cDNA clone was isolated using specific antiserum. The corresponding gene encoding alpha-L-arabinofuranosidase A (abfA) was isolated from a genomic library and cloned into a high copy plasmid vector. By co-transformation of uridine auxotrophic mutants lacking orotidine-5-phosphate decarboxylase activity, the afbA gene was introduced both in A. niger and A. nidulans, using the A. niger pyrA gene as selection marker. The identity of the abfA gene was confirmed by overexpression of the gene product by A. niger and A. nidulans transformants, upon growth using sugar beet pulp as the carbon source.