| Assay Method Information | |
| | NOX Enzyme Inhibition Assay |
| Description: | The inhibition of NADPH oxidase (NOX), which is involved in the generation of reactive oxygen species in red mold, results in the suppression of normal hyphal growth and the formation of abnormal structures known as short-term conidia, thereby inhibiting germination. The abnormal phenotypes caused by the reduction of reactive oxygen species within the red mold can be easily observed through optical microscopy. Through this method, the ability to inhibit the generation of reactive oxygen species by NOX inhibition was evaluated.Specifically, to evaluate the activity of the compounds, a 0.1 mM concentration of the compound was added to a spore suspension of minimal medium diluted to 1/5 (MM20). Spores of the wild-type strain GZ3639 of red mold (105 spores/ml) were inoculated into the medium and incubated at 25° C. or 24 hours, followed by microscopic observation. Compounds that exhibited more than 50% inhibition of germination and short-term conidia formation compared to normal germination were selected for further activity evaluation at concentrations of 50 μM, 25 μM, and 10 μM. Among these, compounds showing 95% germination inhibition at 10 μM were further evaluated at concentrations of 5 μM, 1 μM, 0.5 μM, and 0.1 μM. Based on these results, the concentration of the compound at which the spore germination inhibition rate reached 50% (inhibitory concentration 50%, IC50) was determined. |
| Affinity data for this assay | |
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