Salcha-1 was incubated in six Erlenmeyer flasks (1L) containing 200mL of Potato Dextrose Broth (10g/L Great Value mashed potatoes with 5g/L D-glucose). One of the five flasks received approximately 1.0-1.3g of the following metal compounds: cobalt (II) chloride hexahydrate, cobalt (II) nitrate hexahydrate, cobalt (II) thiocyanate, erbium chloride hexahydrate, or palladium (II) acetate. The sixth flask was reserved as a control and only contained PDB. Salcha-1 was incubated on an Innova 5000 shaker at 135rpm for three weeks at room temperature.
Fungal Compound Extractions from Salcha-1 and Data Analysis
To generate crude extract, approximately 400mL of ethyl acetate was added to each flask to obtain a total volume of 600mL and evaporated …show more content…
Salcha-1, when not incubated with a metal, produced the compound known as maximiscin with a chemical formula of C23H31NO8 with a LC-MS retention time of approximately 7.7 minutes (fig. 3, 9). Salcha-1 also produced a compound around the retention time of 9.2 minutes that was determined to be (Z)-5-(hydroxy(4-hydroxyphenyl)methyl)-3-((E)-1-hydroxy-2,4-dimethyloct-6-enylidene)pyrrolidine-2,4-diene (fig. 1, 9). Both of these compounds were also produced when Salcha-1 was inoculated with either cobalt (II) chloride, cobalt (II) nitrate hexahydrate, erbium chloride hexahydrate. Additionally, Salcha-1 incubated with erbium chloride hexahydrate produced methyl protocatechuate at a retention of 5.1 minutes (fig .2, 7). Salcha-1 with palladium (III) acetate produced a tetramic acid metabolite alone (fig. 8). All three compounds were not produced with incubated with cobalt (II) thiocyanate (fig. 5). All samples produced compounds that were not included in the laboratory database. Due to time constraints and broad search parameters, these compound were not determined if they are novel or already incorporated in the Dictionary of Natural Products. Refer to supplementary materials for inconclusive …show more content…
4, 5). While Astapovich et al. reported gene regulation being altered when exposed to cobalt (II) nitrate and cobalt (II) chloride, maximiscin and the tetramic acid metabolite was produced; this may indicate that gene expression was neither down nor up regulated when compared to the control sample.10 Exposure to erbium chloride also appeared to have minimal effects on Tolypocladium gene regulation and compound production. While the Tolypocladium culture containing erbium chloride produced maximiscin and tetramic acid, methyl protocatechuate was also produced (fig. 7). Methyl protocatechuate is produced when shikimate losses water, and the ability to produce this compound begins to follow the results of Du et al. 6, 11 When maximiscin is produced, it is often accompanied by tetramic acid and shikimate.6 When comparing compound production, palladium acetate and cobalt thiocyanate appeared to have the greatest effect. When co-cultured with palladium acetate, only tetramic acid was produced and blocked the production of maximiscin and shikimate (fig. 8). Cobalt thiocyanate prevented production of all three compounds (fig.
Fungal Compound Extractions from Salcha-1 and Data Analysis
To generate crude extract, approximately 400mL of ethyl acetate was added to each flask to obtain a total volume of 600mL and evaporated …show more content…
Salcha-1, when not incubated with a metal, produced the compound known as maximiscin with a chemical formula of C23H31NO8 with a LC-MS retention time of approximately 7.7 minutes (fig. 3, 9). Salcha-1 also produced a compound around the retention time of 9.2 minutes that was determined to be (Z)-5-(hydroxy(4-hydroxyphenyl)methyl)-3-((E)-1-hydroxy-2,4-dimethyloct-6-enylidene)pyrrolidine-2,4-diene (fig. 1, 9). Both of these compounds were also produced when Salcha-1 was inoculated with either cobalt (II) chloride, cobalt (II) nitrate hexahydrate, erbium chloride hexahydrate. Additionally, Salcha-1 incubated with erbium chloride hexahydrate produced methyl protocatechuate at a retention of 5.1 minutes (fig .2, 7). Salcha-1 with palladium (III) acetate produced a tetramic acid metabolite alone (fig. 8). All three compounds were not produced with incubated with cobalt (II) thiocyanate (fig. 5). All samples produced compounds that were not included in the laboratory database. Due to time constraints and broad search parameters, these compound were not determined if they are novel or already incorporated in the Dictionary of Natural Products. Refer to supplementary materials for inconclusive …show more content…
4, 5). While Astapovich et al. reported gene regulation being altered when exposed to cobalt (II) nitrate and cobalt (II) chloride, maximiscin and the tetramic acid metabolite was produced; this may indicate that gene expression was neither down nor up regulated when compared to the control sample.10 Exposure to erbium chloride also appeared to have minimal effects on Tolypocladium gene regulation and compound production. While the Tolypocladium culture containing erbium chloride produced maximiscin and tetramic acid, methyl protocatechuate was also produced (fig. 7). Methyl protocatechuate is produced when shikimate losses water, and the ability to produce this compound begins to follow the results of Du et al. 6, 11 When maximiscin is produced, it is often accompanied by tetramic acid and shikimate.6 When comparing compound production, palladium acetate and cobalt thiocyanate appeared to have the greatest effect. When co-cultured with palladium acetate, only tetramic acid was produced and blocked the production of maximiscin and shikimate (fig. 8). Cobalt thiocyanate prevented production of all three compounds (fig.