EC Number   |
|---|
   3.1.1.6 | - |
| 3.1.1.6 | 8.6fold, to homogeneity |
| 3.1.1.6 | 85% (NH4)2SO4-saturated bacterial protein extract to anion-exchange chromatography on DEAE-650 (S) column (gradient elution with 0-0.15 M NaCl), EST1 elution at 70 mM NaCl, EST2 elution at 90 mM NaCl, addition of (NH4)2SO4 (25% saturation) to EST1 fraction followed by hydrophobic interaction chromatography on phenyl Sepharose column, purification of main esterase activity (EST1) by elution with 10% (NH4)2SO4-saturated buffer, preparative electrophoresis (native) |
| 3.1.1.6 | after over-expression (up to 20fold activity) by liquid column chromatography, monitored by activity measurements, 300 mg recombinant enzyme from 1 litre culture |
| 3.1.1.6 | ammonium sulfate precipitation and DEAE cellulose column chromatography |
| 3.1.1.6 | at 4°C, precipitation from culture filtrate by 40-80% (NH4)2SO4 saturation followed by anion-exchange (DEAE column, elution with 0-1 M NaCl gradient), t-butyl hydrophobic (elution with 1.7-0 M (NH4)2SO4 gradient) and hydroxyapatite chromatography (elution with 0.02-0.5 M sodium phosphate buffer, pH 6.8), stored at -20°C after addition of 20% glycerol |
| 3.1.1.6 | centrifugation at 4°C, liquid column chromatography |
| 3.1.1.6 | from bacterial extracts by binding to CNBr-activated Sepharose CL6B, 1.5 mg bound protein per ml resin, pH 8.3, 0.1 M sodium hydrogen carbonate, 0.5 M sodium chloride |
| 3.1.1.6 | from bacterial lysate or inclusion bodies by Superose 6 column chromatography in presence of 6-8 M urea for antibody generation, from insect cells by FPLC-based metal affinity chromatography (250 mM imidazole) and anion-exchange chromatography on Q-sepharose column for activity measurements |
| 3.1.1.6 | from the crude commercial enzyme preparation, 27fold |
