Abstract
The co-culture of Trametes versicolor and Ganoderma applanatum is a model of intense basidiomycete interaction, which induces many newly synthesized or highly produced features. Currently, one of the major challenges is an identification of the origin of induced features during the co-culture. Herein, we report a 13C-dynamic labeling analysis used to determine an association of induced features and corresponding fungus even if the identities of metabolites were not available or almost nothing was known of biochemical aspects. After the co-culture of T. versicolor and G. applanatum for 10 days, the mycelium pellets of T. versicolor and G. applanatum were sterilely harvested and then mono-cultured in the liquid medium containing half fresh medium with 13C-labeled glucose as carbon source and half co-cultured supernatants collected on day 10. 13C-labeled metabolome analyzed by LC-MS revealed that 31 induced features including 3-phenyllactic acid and orsellinic acid were isotopically labeled in the mono-culture after the co-culture stimulation. Twenty features were derived from T. versicolor, 6 from G. applanatum, and 5 features were synthesized by both T. versicolor and G. applanatum. 13C-labeling further suggested that 12 features such as previously identified novel xyloside [N-(4-methoxyphenyl)formamide 2-O-beta-D-xyloside] were likely induced through the direct physical interaction of mycelia. Use of molecular network analysis combined with 13C-labeling provided an insight into the link between the generation of structural analogs and producing fungus. Compound 1 with m/z 309.0757, increased 15.4-fold in the co-culture and observed 13C incorporation in the mono-culture of both T. versicolor and G. applanatum, was purified and identified as a phenyl polyketide, 2,5,6-trihydroxy-4, 6-diphenylcyclohex-4-ene-1,3-dione. The biological activity study indicated that this compound has a potential to inhibit cell viability of leukemic cell line U937. The current work sets an important basis for further investigations including novel metabolites discovery and biosynthetic capacity improvement.
Highlights
Secondary metabolites are an important source of valuable drug leads, of which compounds derived from various fungi, especially medicinal fungi of basidiomycetes, represent an important part
Unsupervised principal component analysis (PCA) is well-suited for comparing different biological samples and identifying statistically significant differences (Chen et al, 2007)
Many of the induced features were observed for both samples of T. versicolor and G. applanatum
Summary
Secondary metabolites are an important source of valuable drug leads, of which compounds derived from various fungi, especially medicinal fungi of basidiomycetes, represent an important part. In the case of basidiomycetes, the induction of novel secondary metabolites or enhancement of secreting extracellular enzymes can be achieved by activating cryptic biosynthetic pathways through establishment of a fungi interaction in the co-culture (Peiris et al, 2008; Hiscox et al, 2010). This coculture strategy mimics natural ecosystem, in which interspecies interaction of basidiomycetes is very common (Hiscox et al, 2015, 2017). The co-culture system of Trametes versicolor and Ganoderma applanatum showed an interaction zone, in which the accumulation of a series of known carboxylic acids as well as novel xylosides were observed
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