Table 1.
Properties of the applied enzymes and model reactions. The complexity of the investigated enzymatic reactions ranges from the simple case of cleaving the model substrate ONPG by the enzyme β-Gal up to the cofactor-dependent cascade reaction of BFD and ADH.
| Applied enzyme | β-Gal | BFD | ADH | |
|---|---|---|---|---|
| Substrates | ONPG | Benzaldehyde Acetaldehyde |
Acetophenone | (S)-HPP (cascade intermediate) |
| Products | O-nitrophenol Galactose |
(S)-HPP | (R)-Phenylethanol | (S,S)-PPD |
| Cofactor | None | Magnesium Thiamine disphosphate (both added in flow) |
NADPH (added in flow non-equimolar; regeneration in flow by 2-propanol) |
|
| Mass of monomer [kDa] | 105–106 monomeric enzyme (Tanaka et al., 1975; Maksimainen et al., 2013) |
56 Quaternary structure tetramer (Hasson et al., 1998) |
27 Quaternary structure tetramer (Niefind et al., 2003; Schlieben et al., 2005; Leuchs and Greiner, 2011) |
|
| pH optimum | 4.6 (Tanaka et al., 1975) |
8 (Iding et al., 2000) |
7 for reductions (9 for oxidations) (Riebel, 1996) |
|