In total 86 differentially regulated proteins could be identified by comparing gels from different time points of hypoxic cultivation.
Two critical process parameters were identified, harvest point of the cultivation (harvest) and pH of the extraction buffer (pH), which significantly affected yield (7-fold) and bacterial lysis (35-fold).
In fact, when this pigment was produced in 10 mM glutamine medium at the point of 96 h of cultivation, the intracellular ammonium level of mycelia was reduced to as almost the same as under PP-O production conditions.
CDW corresponding to the end-point of the cultivations was measured in duplicate by filtering a measured volume of the culture through a pre-weighed 0.45 µm pore size nitrocellulose filter (PALL, Michigan, USA).
Thirthy-five stress- and defense-related genes are only up-regulated during the early time point of co-cultivation and thus appear to respond to chemical mediators released by the fungus (cf. Discussion; Figure 5; Figure 6; Additional file 1: Table S1A).
At four time points (2, 4, 6 and 8d) of cultivation, all colonies on each CBM plate were collected, resuspended in 5 mL liquid CGM medium and quantified based on A600.
In these cultures, maximum TAG production of 2.3 (±0.3) g L−1, representing 40.4 (±3.1) % of CDW occurred after 6 days of cultivation, at which point the residual glycerol was completely consumed.
However, the depletion of nitrogen and the initiation of simultaneous utilization of xylose and glucose in the solution occurred between 4 days and 5 days, and fatty acid production of 11.0 (±1.3) g L-1 corresponding to 45.8 (±1.3) % of the cell dry weight was obtained after 7 days of cultivation, at which point the residual sugars were consumed.
Finally, two genes encoding probable extracytoplasmic solute receptors (H16_A2779, H16_A0337; two- to fourfold each) exhibited increased expression levels at the first three time points during cultivation in presence of DTDP.
From his point of view, the cultivation of oneself leads to the freedom of thought, freedom to act, freedom to assert oneself as an individual, freedom to access knowledge, and freedom to determine one¿s own role in society.
