Table 2.
PA3944 enzyme | substrate | KM (mM) | kcat(s−1) | kcat/KM(M−1 s−1) |
---|---|---|---|---|
tagged | polymyxin B (constant [AcCoA])a | 2.57 ± 0.23 | 0.653 | 2.54 × 102 |
colistin (constant [AcCoA])a | 2.28 ± 0.17 | 0.672 | 2.95 × 102 | |
DAB (constant [AcCoA])a,b | 8.60 ± 1.59 | 0.264 | 30.7 × 10 | |
AcCoA (constant [polymyxin B])c | 0.118 ± 0.028 | 0.602 | 5.10 × 103 | |
AcCoA (constant [colistin])c | 0.105 ± 0.017 | 0.471 | 4.49 × 103 | |
cleaved | polymyxin B (constant [AcCoA])a | 4.00 ± 0.32 | 0.923 | 3.65 × 102 |
colistin (constant [AcCoA])a | 2.50 ± 0.13 | 0.894 | 3.58 × 102 | |
DAB (constant [AcCoA])a,b | 10.0 ± 1.2 | 0.391 | 3.91 × 10 | |
AcCoA (constant [polymyxin B])c | 0.137 ± 0.020 | 0.610 | 4.45 × 103 | |
AcCoA (constant [colistin] )c | 0.118 ± 0.021 | 0.456 | 3.86 × 103 |
The concentration of AcCoA was held constant at 0.5 mM while that of the acceptor substrate(s) was varied.
At DAB concentrations of >10 mM, substrate inhibition was observed; therefore, kinetic parameters were calculated on the basis of data collected from 0 to 10 mM DAB.
The concentration of polymyxin B or colistin was held constant at 5 mM while that of AcCoA was varied.