1GK0
Structure-based prediction of modifications in glutarylamidase to allow single-step enzymatic production of 7-aminocephalosporanic acid from cephalosporin C
Experimental procedure
| Experimental method | MAD |
| Source type | SYNCHROTRON |
| Source details | ESRF BEAMLINE BM14 |
| Synchrotron site | ESRF |
| Beamline | BM14 |
| Temperature [K] | 100 |
| Detector technology | IMAGE PLATE |
| Detector | MAR scanner 345 mm plate |
| Wavelength(s) | 0.8854,0.9776,0.9779 |
| Spacegroup name | C 1 2 1 |
| Unit cell lengths | 228.366, 69.910, 113.550 |
| Unit cell angles | 90.00, 97.57, 90.00 |
Refinement procedure
| Resolution | 20.000 * - 2.500 |
| R-factor | 0.207 |
| Rwork | 0.207 |
| R-free | 0.22200 |
| Structure solution method | MAD |
| RMSD bond length | 0.007 |
| RMSD bond angle | 24.300 * |
| Data reduction software | XDS |
| Data scaling software | XDS |
| Phasing software | CNS |
| Refinement software | CNS (1.0) |
Data quality characteristics
| Overall | Outer shell | |
| Low resolution limit [Å] | 20.000 | 2.600 |
| High resolution limit [Å] | 2.500 | 2.500 |
| Rmerge | 0.045 | 0.095 |
| Number of reflections | 118686 | |
| Completeness [%] | 96.8 | 79.4 |
| Redundancy | 2.05 | 1.63 |
Crystallization Conditions
| crystal ID | method | pH | temperature | details |
| 1 | VAPOR DIFFUSION, HANGING DROP | 7.5 * | 18 * | CRYSTALS GROWN AT 18 C, HANGING DROP PRECIPITATION AGENT: 1.5-2.0 M POTASSIUM PHOSPHATE PH:7.0 -9.0, pH 8.00 |
Crystallization Reagents in Literatures
| ID | crystal ID | solution | reagent name | concentration (unit) | details |
| 1 | 1 | drop | protein | 20-30 (mg/ml) | |
| 2 | 1 | drop | potassium phosphate | 0.5 (M) | pH7.5 |
| 3 | 1 | drop | dithiothreitol | 5 (mM) | |
| 4 | 1 | reservoir | potassium phosphate | 1.5-2.0 | pH7.0-9.0 |
