Table 5 Details of assay reactions for determination of enzyme activity and inhibition assays in the presence of various dilutions of P. sidoides extract
Enzyme | Enzyme Activity Assay reaction mixtures | P. sidoides inhibition Assay reaction mixtures | Incubation time |
|---|---|---|---|
NDK | 100 mM K-PO4 buffer (pH 6.8), 250 mM KCl 5 nM enzyme, 0.2 M Thymidine diphosphate | 100 mM K-PO4 buffer (pH 6.8), 250 mM KCl 5 nM enzyme, 0.2 M Thymidine diphosphate | 40 mins |
HSK | 50 mM HEPES buffer (pH 7.0), 450 mM KCl 704 nM enzyme, 10 mM Homoserine | 50 mM HEPES buffer (pH 7.0), 450 mM KCl 704 nM enzyme, 10 mM Homoserine | 4 h |
AK | 100 mM Tris buffer (pH 7.0), 250 mM KCl 223 nM enzyme, 10 mM Na-acetate | 100 mM Tris buffer (pH 7.0), 250 mM KCl 223 nM enzyme, 10 mM Na-acetate | 24 h |
GK | 100 mM Tris buffer (pH 7.0), 250 mM KCl 208.6 nM enzyme, 100 mM Glycerol | 100 mM Tris buffer (pH 7.0), 250 mM KCl 208.6 nM enzyme, 100 mM Glycerol | 24 h |
ThiL | 100 mM Tris buffer (pH 8.0), 250 mM KCl 2074 nM enzyme, 1 mM Thiamine monophosphate | 100 mM Tris buffer (pH 8.0), 250 mM KCl 2074 nM enzyme, 1 mM Thiamine monophosphate | 5 h |
RBKS | 100 mM Tris buffer (pH 7.2), 100 mM KCl 250 nM enzyme, 10 mM d-ribose | 100 mM Tris buffer (pH 7.2), 100 mM KCl 250 nM enzyme, 10 mM d-ribose | 4 h |
AsK | 100 mM Tris-HCl buffer (pH 7.5), 178.2 nM enzyme 10 mM l-Aspartic acid | 100 mM Tris-HCl buffer (pH 7.5), 178.2 nM enzyme 10 mM l-Aspartic acid | 6 h |
SK | 100 mM K-PO4 buffer (pH 6.8), 500 mM KCl 10 nM enzyme, 8 mM shikimic acid | 100 mM K-PO4 buffer (pH 6.8), 500 mM KCl 10 nM enzyme, 8 mM shikimic acid | 20 mins |