3.4 Chemical characteristics of the color fractions ofPenicillium gravinicasei P3SO332, Clonostachys roseaP2SO329 and Penicillium gravinicasei P3SO332
Fermentation was carried out in 125 mL Erlenmeyer flasks containing 50 mL of broth medium with the different sources of carbon or nitrogen that were selected in the previous results. This procedure was repeated 20 times and resulted in 1,000 mL of fungal cultivation. Broth containing plugs of mycelium was mixed with ethyl acetate (1:1 v/v) and shaken in an orbital shaker (100 rpm for 24 h). The extraction produced colorant yields of 105 mg/L by Penicillium sclerotiorum P3SO224, 83 mg/L by Clonostachys rosea P2SO329 and 142 mg/L byPenicillium gravinicasei P3SO332, and their metabolites were analyzed using LC/MS-MS.
The EtOAc extract of P. sclerotiorum P3SO224 showed a main peak eluting at 15 min (See Supplementary Material) and a molecular ion of m/z 391 [M+H]+ with competitive losses of -42 (C2H2O) and -60 Da C2H4O2), related to neutral losses on the acetyl group. Moreover, after the loss of 60 Da, a carbon monoxide loss (- 28 Da) yielding the ion m/z 303. These characteristics allowed us to propose that the this compound is the chlorinated azaphilone named sclerotiorin (Peres et al. 2023) (Fig. 4,1 ).
The EtOAc extract of the fungus Clonostachys rosea P2SO329 showed four main chromatographic peaks eluting at 3.6, 5.5, 7.8 and 8.5 min. From these, only the peak eluting at 5.5 min was possible to annotate. The positive and negative modes displayed m/z of 171 [M+H]+ and 169 [M-H]-, both with consecutive losses of - 18 Da (water), - 15 Da (methyl) and – 28 Da (CO), all consistent with the annotation of the compound is penicillic acid ( Fig. 4, 2) [30] .