niger (predicted) proteins. One protein (6715) that

did not match an A. niger protein, probably because it was missed or truncated during sequencing, had a significant match to a protein from N. crassa [UniProt: NCU04657]. Only 6 proteins (8 spots) were identified as proteins in the Swiss-Prot database and thus regarded as fully characterised. Otherwise, the proteins were registered in the NCBInr database as it contains the protein entries predicted from the sequencing of the A. niger CBS 513.88 genome [22]. Per primo March 2009 the predicted proteome based on this sequencing Z-IETD-FMK nmr project contained 13906 predicted proteins of which 47.1% had automatically assigned GO annotations and only 154 proteins had been assigned as manually reviewed in the UniProtKB database [39]. To circumvent the limited number of annotated proteins, we assigned annotations based on sequence similarity to characterised Swiss-Prot proteins in other species using BlastP [40]. A protein annotation was assigned to a protein if it had more than 80% sequence identity to a characterised Swiss-Prot protein and a “”putative”" annotation to proteins that had 50-80% sequence identity to a characterised protein. Other proteins were assigned a “”predicted”" function if InterPro domains were predicted using InterProScan [41]. In this way, the identified proteins consisted of 6 (8 spots) fully characterised, 12 with annotation based on sequence

similarity, 19 with putative annotation, 13 with predicted function and 6 (7 spots) uncharacterised proteins. The proteins with known functions were mainly old involved in AZD0156 order processes as: polysaccharide degradation; carbon-, nitrogen- and amino acid metabolism; energy production; protein synthesis, folding and degradation; redox balance and protection

against oxidative stress. None of the characterised proteins were known to participate in secondary metabolite biosynthesis. A fatty acid synthase subunit alpha [UniProt: A2Q7B6] was identified, which was present at higher levels on SL compared to on S and L (cl. 35). This protein may contribute to fatty acid biosynthesis to be incorporated in the cell LY2835219 cell line membrane; however it may also be an unrecognised polyketide synthase. One gene coding for a predicted aldo/keto reductase [UniProt: A2Q981] was located adjacent to the predicted FB2 biosynthesis cluster in the A. niger genome. But this protein was present at higher levels on starch-containing media (cl. 3) and therefore did not correlate with FB2 production. Furthermore, proteins involved in secondary metabolite synthesis or processes associated with transport or self-protection are not necessarily located within the clusters. One example is a reductase found to participate in aflatoxin biosynthesis in A. parasiticus, although it is not located within the aflatoxin cluster and was regulated differently than the aflatoxin cluster genes [42].