Table 1.
Purification of Chi18H8 (A) from E. coli BL21 StarTM (DE3)/pET24b(+)::chi18H8 IBs and 53D1 (B) from S. coelicolor/pIJ86::53D1 culture broth.
| Purification step | Volume (mL) | Total proteins (mg) | Total activity (U) | Specific activity (U/mg protein) | Purification (-fold) | Yield (%) |
|---|---|---|---|---|---|---|
| (A) | ||||||
| IBs | 10.6 | 123.0 | 33.4 | 0.27 | 1.0 | 100.0 |
| Soluble fraction from IBs | 53.0 | 71.8 | 2623.0 | 36.7 | 78.5 | 84.0 |
| (B) | ||||||
| Crude broth | 1000.0 | 1125.0 | 1080.0 | 0.96 | 1.0 | 100.0 |
| Ammonium sulfate precipitation | 200.0 | 1090.0 | 1067.1 | 0.98 | 1.0 | 98.8 |
| Affinity chromatography | 114.1 | 34.9 | 956.6 | 27.4 | 28.5 | 88.5 |
For both proteins, data are relative to cells (Chi18H8) or cell-free culture broth (53D1) from 1 L of culture. Activity was assayed on 4-MU-(GlcNAc)2 as substrate, in 100 mM sodium acetate pH 5.0.