Table 1.
Evaluating ACP Sfp/AcpH reaction efficiency
| a | E.coli ACP | |||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| apo #1 | butanoyl | apo #2 | apo #1 | octanoyl | apo #2 | apo #1 | rhodamine | apo #2 | apo #1 | coumarin | apo #2 | |
| mass (mg) | 1.9 | 1.7 | 1.8† | 1.6 | 1.2 | 1.0 | 1.6 | - | 1.0 | 1.6 | - | 1.4 |
| step yield | - | 92% | 99% | - | 77% | 83% | - | - | 64% | - | - | 85% |
| cycle yield | 98% | 64% | 64% | 85% | ||||||||
| b | MBP-PaACP | GFP-PaACP | Luciferase-ACP | |||||||||
| apo #1 | rhodamine | apo #2 | apo #1 | rhodamine | apo #2 | apo #1 | rhodamine | apo #2 | ||||
| mass (mg) | 1.7 | 1.3 | 0.9 | 2.5 | 2.2 | 1.6 | 1.7 | 1.5 | 1.0 | |||
| step yield | - | 92% | 74% | - | 86% | 75% | - | 82% | 68% | |||
| cycle yield | 57% | 65% | 56% | |||||||||
All free (d) and fusion (e) ACPs were labeled and unlabeled in order to validate reaction yields.
†
Elevated yield calculation for obtaining apo-ACP from butanoyl-ACP is most likely due to minor protein or buffer contamination for this sample. Reaction completion was monitored by Urea-PAGE (Supplementary Fig. 16) and SDS-PAGE (Supplementary Fig. 17) as appropriate.