Molecular basis of glucoamylase overproduction by a mutagenised industrial strain of Aspergillus niger.

We have compared a mutagenized strain of Aspergillus niger (S1), used industrially for glucoamylase production, and a related low glucoamylase-producing strain (S2) with a laboratory strain of A. niger (AB4.1). Our aim was to assess the properties of S1 in relation to the laboratory strain and to account at the molecular level for the basis of its glucoamylase overproduction. Both S1 and S2 have similar multiple copies of the glucoamylase-encoding gene (glaA) but only S1 has enhanced glaA transcript and glucoamylase levels compared to AB4.1 that has a single copy of the glaA gene. Glucoamylase production by S1 and AB4.1 was repressed by xylose and induced by starch but, in S2, remained unaffected by carbon source. S1 also secreted elevated levels of alpha-amylase relative to both S2 and AB4.1 but the production of alpha-glucosidase was low in all three strains. The gene encoding aspergillopepsin (pepA), an abundant secreted aspartyl protease, was present as a single copy in all strains but no aspergillopepsin could be detected by Western blotting in either S1 or S2 culture supernatants. We conclude that A. niger strain improvement by mutagenesis and screening for glucoamylase overproduction has led to glaA gene multiplication and an expression defect in the pepA gene.