ture ?l trates were performed. Neither www.selleckchem.com/products/MLN8237.html chitin, glucan nor man nan active hydrolases were detected in the culture broth during exponential growth. In agreement to its high transcript levels during carbon starvation, NagA was the most abundant extracellular hydro lase involved in chitin degradation at day 1, 3 and 6. However, the chitinase ChiB was, in contrast to its strong transcriptional upregulation, only marginally detected in ?ltrates at day 1. Both observations correspond well to the presence and absence of predicted signal peptide sequences for NagA and ChiB, respectively. Interestingly, ChiB of A. niger showed only low extracellular abundance, whereas the A. nidulans ChiB was identi?ed as the major extracellular autolytic chitinase during carbon starvation. The absence of ChiB in the culture broth of A.

niger could explain why hyphal ghosts remained intact but were reported to fragment in aging cultures of A. nidulans. In concordance with its expression pro ?le, the glucanase AgnB was detected extracellularly at day 1 and 3. While GelD was the only reliably detected B glucanase during expo nential growth, various B glucanases with predicted sig nal peptide sequences were detected at day 1, 3 and 6 of carbon starvation. Among the predicted mannanases, only An04g09650 was reliably detected in ?ltrates at later time points. In agreement with increasing extracellular protease activity and expression pro?les, a number of proteases with predicted signal peptide sequences were identi ?ed in culture ?ltrates of day 1, 3 and 6. Among them, PepA, the major extracellular pro tease, was most abundant.

However, although PepB has a predicted signal peptide sequence and showed strong transcriptional induction, it was not detected in culture ?ltrates. Transcriptionally induced proteases lacking predicted signal peptide sequences were not detected in culture ?ltrates, with the only excep tion of An01g00370. Similar results have been previ ously reported for A. niger by Braaksma et al. who proposed that the high extracellular abundance of An01g00370 is likely a result of non classical secretion rather than lysis. The secretome during starvation conditions was clearly enriched by an additional group of proteins with strong similarity to phospholipases. Together the four putative phospholipases, An16g01880, An09g02180, An01g14940 and An02g13220 constituted on average about 7% of all detected extracellular proteins during day 1, 3 and 6.

All except An02g13220 were transcriptionally induced during carbon starvation. This high abundance of pre dicted phospholipases during carbon starvation might be indicative for a role of membrane lipids as alternative car bon source during secondary growth. The Carfilzomib complete list of identi?ed extracellular proteins is given in selleck compound Additional ?le 7. Discussion The present study is the ?rst system wide description of the carbon starvation response in a ?lamentous fun gus. The application of bioreactor technology allowed highly reproducib