Apple, banana, and grape fruits no longer suitable for consumption
were ground in a blender (Waring commercial, McConnellsburg) to obtain
particles of 1–2 mm of diameter. The whole digestate was thawed
and manually mixed with shredded fruits in a ratio of 70:30 w/w. The
substrate composition is reported inTable 1 . Wood sawdust, a local carpentry byproduct,
was added to the whole digestate to reduce the free water and adjust
the moisture content to 73%. SSF was carried out in micropropagation
boxes equipped with a 0.45 μm filter (Microbox, Micropoli, Italy),
each containing 70 g of substrate. Filled boxes were subjected to
two consecutive cycles of sterilization (121 °C for 15 min each).
For the analyses, seven time points were considered: T0, T1, T2, T3,
T4, T5, and T6 corresponding to 0, 3, 6, 13, 20, 27, and 34 days after
inoculation. T. reesei RUT-C30 and T. atroviride Ta13 were routinely grown on slants
of PDA at 26 °C. Then, 100 μL of conidia suspension (106 in sterile distilled water) was inoculated in each box; four
replicate boxes were produced for each time point and for each strain.
In addition, four replicate boxes were not inoculated and served as
a control (SSF-NI). All of the boxes were incubated at 26 °C
and 60% relative humidity (RH) under illumination of 12 h light/12
h dark cycles, using daylight tubes 24 W/m2, in a climatic
chamber (model 720, Binder) for 34 days. For each time point, and
for each strain, one replicate was used for fungal biomass quantification
and stored at −80 °C until use. The remaining three replicates
were treated as described below to obtain the crude extract for enzymatic
and metabolomic analyses.
were ground in a blender (Waring commercial, McConnellsburg) to obtain
particles of 1–2 mm of diameter. The whole digestate was thawed
and manually mixed with shredded fruits in a ratio of 70:30 w/w. The
substrate composition is reported in
was added to the whole digestate to reduce the free water and adjust
the moisture content to 73%. SSF was carried out in micropropagation
boxes equipped with a 0.45 μm filter (Microbox, Micropoli, Italy),
each containing 70 g of substrate. Filled boxes were subjected to
two consecutive cycles of sterilization (121 °C for 15 min each).
For the analyses, seven time points were considered: T0, T1, T2, T3,
T4, T5, and T6 corresponding to 0, 3, 6, 13, 20, 27, and 34 days after
inoculation. T. reesei RUT-C30 and T. atroviride Ta13 were routinely grown on slants
of PDA at 26 °C. Then, 100 μL of conidia suspension (106 in sterile distilled water) was inoculated in each box; four
replicate boxes were produced for each time point and for each strain.
In addition, four replicate boxes were not inoculated and served as
a control (SSF-NI). All of the boxes were incubated at 26 °C
and 60% relative humidity (RH) under illumination of 12 h light/12
h dark cycles, using daylight tubes 24 W/m2, in a climatic
chamber (model 720, Binder) for 34 days. For each time point, and
for each strain, one replicate was used for fungal biomass quantification
and stored at −80 °C until use. The remaining three replicates
were treated as described below to obtain the crude extract for enzymatic
and metabolomic analyses.
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