Identification of a Novel Biosynthetic Gene Cluster in Aspergillus niger Using Comparative Genomics

Previously, DNA microarrays analysis showed that, in co-culture with Bacillus subtilis, a biosynthetic gene cluster anchored with a nonribosomal peptides synthetase of Aspergillus niger is downregulated. Based on phylogenetic and synteny analyses, we show here that this gene cluster, NRRL3_00036-NRRL3_00042, comprises genes predicted to encode a nonribosomal peptides synthetase, a FAD-binding domain-containing protein, an uncharacterized protein, a transporter, a cytochrome P450 protein, a NAD(P)-binding domain-containing protein and a transcription factor. We overexpressed the in-cluster transcription factor gene NRRL3_00042. The overexpression strain, NRRL3_00042(OE), displays reduced growth rate and production of a yellow pigment, which by mass spectrometric analysis corresponds to two compounds with masses of 409.1384 and 425.1331. We deleted the gene encoding the NRRL3_00036 nonribosomal peptides synthetase in the NRRL3_00042(OE) strain. The resulting strain reverted to the wild-type phenotype. These results suggest that the biosynthetic gene cluster anchored by the NRRL3_00036 nonribosomal peptides synthetase gene is regulated by the in-cluster transcriptional regulator gene NRRL3_00042, and that it is involved in the production of two previously uncharacterized compounds.

Automated summary

The study found that a biosynthetic gene cluster in Aspergillus niger is downregulated in co-culture with Bacillus subtilis. Gene analysis revealed that the cluster contains various genes including nonribosomal peptides synthetase and transcription factor. Overexpression of the transcription factor gene led to reduced growth rate and production of two previously unknown compounds, which were reverted to wild-type phenotype upon deletion of the nonribosomal peptides synthetase gene. This suggests that the biosynthetic gene cluster is regulated by the transcriptional regulator gene and is involved in the production of these compounds.