Table I.
Steady-state kinetic parameters for various ncAAs and PylRS enzymes.
| Organism | ncAA |
Km [mmol.dm−3] |
kcat [1/s] |
Assaya | Enzyme variantb | Structurec | Ref |
|---|---|---|---|---|---|---|---|
| D. hafniense | l-pyrrolysine | 0.044 | 0.325 | 1 | PylSc | - | [6] |
| M. barkeri | Nε-((R)-tetrahydrofuran-2-carbonyl)-l-lysine | 0.39 | 0.062 | 1 | WT | - | [66] |
| Nε-(cyclopentanecarbonyl)-l-lysine | 5.6 | 0.04 | 1 | WT | - | ||
| l-pyrrolysine | 0.055 0.02 |
0.105 0.003 |
1 2 |
WT WT |
- 2Q7H, 2ZIM, 2ZCE |
[66] | |
| Nε-((cyclopentyloxy)carbonyl)-l-lysine | 0.55 | 0.015 | 1 | WT | 2Q7G | [66] | |
| l-pyrrolysine | 0.053 n.d. n.d. |
n.d. 0.1 0.24 |
1 1 1 |
WT WT Δ92PylS |
2Q7H, 2ZIM, 2ZCE 2Q7H, 2ZIM - |
[23] [6] |
|
| Nε-((cyclopentyloxy)carbonyl)-l-lysine | 0.67 | n.d. | 1 | WT | 2Q7G | [23] | |
| Nε-acetyl-l-lysine | 22.3 | 0.0341 | 1 | AcKRS3 | - | [36] | |
| Nε-d-prolyl-l-lysine | 0.5 | n.d. | 1 | WT | - | [23] | |
| M. mazei | 3-iodo-l-phenylalanine | 0.44 | 0.0044 | 2 | IFRS | 4TQD | [22b] |
| l-phenylalanine | 21 14 22.8 |
0.037 0.062 0.075 |
1 1 1 |
FRS1 FRS2 FRS3 |
- - - |
[39a] |
|
| l-pyrrolysine | 0.02 | 0.0083 | 2 | WT | 2Q7H, 2ZIM, 2ZCE | [22b] | |
| Nε-acetyl-l-lysine | 35.3 7.8 |
0.0323 0.00731 |
1 1 |
AcKRS1 AcKRS2 |
4Q6G - |
[36] |
a
(1) - ATP-[32P]PPi exchange
(2) - aminoacylation
b
AcKRS1- M. mazei PylRS variant L301M/Y306L/L309A/C348F; AcKRS2- M. mazei PylRS variant L301M/L305M/Y306M/C348S; AcKRS3- M. barkeri PylRS variant L266M/L270I/Y271F/L274A/C313F; FRS1- M. mazei PylRS variant N346A/C348L; FRS2- M. mazei PylRS variant A302L/Y306M/N346S/C348L/Y384L; FRS3- M. mazei PylRS variant A302F/Y306L/N346T/C348F/Y384L; IFRS- M. mazei PylRS variant N346S/C348I
c
All structures are of M. mazei enzymes.