From single cultures of bacterial isolates and fungus/bacteria co-cultures on agar, 24 different compounds could be identified by comparing the HPLC-MS profiles of the respective agar extracts with an in-house HPLC-UV–VIS database (Table 1). The mix of the different exudates was to some degree isolate-specific. Multi dimensional statistical (MDS) data analysis illustrates which individual cultures and co-cultures form clusters, and which cultures could be considered similar to one another, on the basis of patterns and combinations due to the presence or absence of exudate compounds.
This approach indicates that the inhibition of the fungus in co-culture (Figure 3; MW2, 4, 9; M2, 4, 5) was dependent on the presence of compounds of two groups (Figure 4; Table 2). These are group Fludarabine mouse 1, made up by compounds 1, 2, 3 and sometimes 4 (Figure 4; □), and group 2, consisting of compounds
16, 17, and 18 (Figure 4; ◊), each enclosed by circles. Group 1 consists of a ß-carboline see more alkaloid usually extracted from Actinomycetes (1-acetyl-β-carboline, 1 in Table 1), containing an indole tricyclic ring and is cytotoxic, anti-microbial and an enzyme inhibitor [31]. The other three metabolites in this group are polyene macrolide antibiotics, containing a lactose ring and act against ergosterol of fungal membranes. Filipin is more toxic than lagosin and all three cause excess leakage of K [32]. Group 2 consist of a peptide antibiotic (stenothricin, 16) that affects glycolytic and lipolytic proteins, and inhibits cell wall formation [33]. The other two compounds (17, 18) are auxins or auxin antagonists (plant
hormone derivatives) and may affect many aspects of plant growth and development [34]. Compounds 17 and 18 were generally not released or present from single cultures of either bacteria or fungus, and this is consistent with Rutecarpine their roles more directly in plants. Two other well separated metabolites are worth mentioning (i.e. Figure 4/Table 1, 13 and 24). Thiolutin (Δ) is a well studied broad spectrum indole alkaloid which inhibits energy metabolism, RNA synthesis (RNA polymerase), glucose metabolism and carbon use [35]. N-hydroxy phenyl acetic acid methyl ester is a derivative of indole propionic acid and is a weak alkaloid and anti-microbial compound, acting mainly against Gram-negative bacteria [34]. Most effective in the inhibition of fungal growth are combinations and the presence of compounds belonging to both group 1 and group 2, however, not all metabolites included in these groups are apparently necessary for inhibition. Table 1 Compilation of compounds identified by HPLC-MS from exudates released into the agar by the different streptomycte isolates, singly or in co-culture with N. parvum Number Compound Number Compound 1 1-Acetyl-β-carboline 13 Thiolutin 2 Lagosin 14 NL 19 KF RT 3.